Piezoelectric materials have emerged as promising electroactive biomaterials in regenerative medicine owing to their ability to convert mechanical forces into electrical signals and vice versa. These materials reproduce aspects of the body's native bioelectric microenvironment and influence key cellular processes, including adhesion, proliferation, migration, and differentiation. Clinically, piezoelectricity has been exploited, in dental implants, where electromechanical activity enhances osseointegration and long-term stability. This review provides a comprehensive overview of the principles of piezoelectricity, the major classes of piezoelectric materials, and recent advances in fabrication strategies such as electrospinning, additive manufacturing, and nanogenerators. Applications across bone, nerve, cartilage, skin, and cardiovascular tissues are critically examined, with emphasis on mechanosensitive ion channels, intracellular signalling pathways, and gene regulation. Safety concerns, including ion release from ceramic materials, and the emergence of biocompatible, lead-free alternatives are discussed alongside translational barriers related to scalability, regulatory approval, and device integration. The aim of this review is to provide a mechanistic and clinically oriented perspective that informs the design of next-generation piezoelectric materials. Finally, future directions in self-powered implants and piezoelectric catalysis are highlighted to support their clinical translation for tissue repair and regenerative therapies. STATEMENT OF SIGNIFICANCE: This review uniquely integrates the fundamental and translational aspects of piezoelectric biomaterials in regenerative medicine. We highlight how piezoelectric cues regulate cell behaviour through electrical stimulation, ion channel activation, particularly Ca²⁺ flux and downstream signalling pathways such as Wnt/GSK3β and PI3K/Akt. By linking these mechanisms to gene expression profiles and functional outcomes across bone, nerve, cartilage, cardiovascular, and skin tissues, this work provides a tissue-specific perspective that has not been comprehensively addressed before. Importantly, we emphasise the multifunctionality of piezoelectric scaffolds, showcasing their immunomodulatory, angiogenic, and biomechanical benefits. The review further bridges insights across chemistry, biology, materials science, and biofabrication, offering constructive guidance for designing next-generation, clinically translatable piezoelectric biomaterials.
Hard tissues are core components maintaining the mechanical integrity and physiological functions of organisms but have limited self-repair capacity. Traditional clinical strategies for hard tissue healing or regeneration face challenges, including donor scarcity. As engineered 3D cell culture systems that recapitulate tissue/organ-specific structures and functions, organoids outperform conventional 2D cell culture systems and animal models, thereby holding substantial potential for hard tissue healing or regeneration. Regrettably, constructing desired hard tissue organoids is highly demanding, as simulating the intricate structures and mechanical properties of native hard tissues is inherently challenging. Recently, hard tissue organoids have gradually transformed from a conceptual framework into tangible research objects. Focusing on this challenging yet promising field, this review comprehensively delineates the desired features and corresponding viewpoints of hard tissue organoids. It then systematically summarizes the design and construction strategies for organoids, the specific advances in hard tissue organoid development, and their diverse biomedical applications. Furthermore, contemporary challenges and prospective directions in the development and translational application of hard tissue organoids are thoroughly discussed. The summarized insights and proposed perspectives aim to guide the design and engineering of next-generation, or even ideal hard tissue organoids. This review may further facilitate the widespread application and efficient use of hard tissue organoids, as well as related advanced matrix technologies, in drug screening and hard tissue regeneration. STATEMENT OF SIGNIFICANCE: As engineered 3D systems recapitulating tissue/organ-specific structures and functions, organoids outperform conventional 2D cell cultures and animal models. Regrettably, constructing desired hard tissue organoids remains demanding, as simulating the intricate structures and superior mechanical properties of native hard tissues is inherently challenging. Focused on this promising field, this review comprehensively outlines the desired features and corresponding viewpoints of hard tissue organoids. It also summarizes the design and construction strategies for organoids, the latest advances in hard tissue organoid development, and their diverse biomedical applications. Finally, current challenges and prospective directions in their development and translational application are thoroughly discussed, with a focus on multidimensional biomimicry, simulation of typical gradient features, and integrated assembly of adjacent tissue organoids.
Critical-sized bone defects caused by trauma, tumor resection, injury, and/or surgical intervention are posing significant clinical challenges. Bone tissue regeneration is crucial for restoring critical-sized bone defects. Central to the bone regenerative capability is the dynamic interplay between bone cells, particularly osteocytes, which are the most abundant and long-lived bone cells, functioning as key mechanosensors in bone. Osteocytes detect mechanical stimuli, for example, fluid shear stress, compressive or tensile strain, and hydrostatic pressure, and convert these into biochemical signals through mechanotransduction. The biochemical signals (eg, calcium ions, Wnt, etc.) regulate osteoblast and osteoclast-mediated remodeling. Osteocytes communicate with osteoblasts and osteoclasts via paracrine factors, including nitric oxide, prostaglandins, and sclerostin. Moreover, estrogen deficiency is known to alter osteocyte mechanosensitivity, impair osteocyte signaling, and dysregulate bone remodeling. Understanding how mechanical and hormonal factors affect osteocyte signaling is essential for developing effective therapeutic interventions. This concise review explores the role of osteocyte mechanosensing and mechanotransduction in bone tissue regeneration to improve bone healing, especially in critical-sized bone defects. The cellular and molecular mechanisms underlying bone regeneration and remodeling are discussed, including the role of stem cells in bone regeneration, that is, osteogenic differentiation potential and secretion of bioactive factors that promote new bone formation and vascularization. Finally, we explore the translational and clinical implications of osteocyte mechanobiology, discussing current challenges and potential advancements in bone tissue engineering and regenerative medicine. By integrating fundamental mechanobiological principles with clinical strategies, this concise review highlights the clinical potential of modulating osteocyte behavior for improved bone regeneration.
The global shortage of donor organs and the limitations of conventional in vitro models stress the urgent need for advanced liver tissue engineering and regenerative medicine strategies. These approaches aim to create physiologically relevant platforms for drug testing and develop transplantable tissues to restore liver function. Decellularization offers unique advantages by providing native extracellular matrix architecture and biochemical cues that support cell adhesion, differentiation, and vascularization. Complementary technologies such as three-dimensional (3D) bioprinting and microfluidics enable precise spatial organization of multiple cell types and dynamic perfusion, improving tissue functionality and disease modeling. Together, these innovations facilitate the development of high-fidelity liver constructs and organ-on-chip systems for studying pathologies like fibrosis and steatosis, as well as for preclinical drug screening. In this review we summarize current methods for liver decellularization and explore its role as a regenerative medicine strategy. We also examine applications in disease modeling, with emphasis on 3D bioprinting and microfluidic platforms, and discusses emerging vascularization techniques. Collectively, these insights highlight the progress and remaining challenges in engineering functional liver tissues for clinical and research applications.
Collagen plays a critical role in tissue engineering and regenerative medicine due to its biocompatibility and structural features. Identifying an optimal extraction method that preserves both functional and structural integrity remains challenging. This study aimed to compare eleven collagen extraction protocols to determine the most effective method for producing high-quality collagen. In this a comparative experimental study, collagen was extracted using eleven protocols and characterized using scanning electron microscope (SEM), Fourier-transform infrared spectroscopy (FTIR), XRD, Raman spectroscopy, MTT and anti-inflammatory assays, lactate dehydrogenase (LDH) cytotoxicity, total antioxidant capacity (TAC), blood compatibility tests, protein quantification (BCA and Bradford), sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and solubility evaluations at varying pH and salt concentrations. The most promising sample was further evaluated In vivo via subcutaneous transplantation in mice, followed by histological analysis at 30 days. Among the evaluated protocols, protocol 4 consistently exhibited superior performance, yielding approximately 35 mg collagen per g tissue. SEM, FTIR, XRD, and Raman analyses confirmed preserved triple-helical structure and fiber morphology. Protocol 4 showed high cell viability, low cytotoxicity, optimal blood compatibility, significant antioxidant activity, and improved protein content (P<0.05). In vivo studies demonstrated normal tissue regeneration, including collagen deposition, angiogenesis, and epidermal restoration, without inflammatory response. Other samples exhibited incomplete structural features, reduced yield, or increased cytotoxicity. Protocol 4 provides a robust approach for extracting collagen with high structural integrity, functional stability, and excellent biocompatibility. This optimized collagen may be suitable for biomedical applications and could support further translational research for collagen-based therapeutic products.
Electrospinning is a flexible and viable technique for producing ultrathin fibers. Over the past decade, remarkable progress has been made in the development of electrospinning techniques to fabricate customized nanofibers (NFs) that fulfill specific application needs. This study provides a comprehensive overview of advanced electrospinning approaches, such as coaxial, aligned, template-assisted electrospinning (TAE), near-field electrospinning (NFE), and layer-by-layer (LBL) fabrication, used for the production of three-dimensional (3D) NFs scaffolds. Recently, multilayer (ML) 3D NFs scaffolds have gained immense attention due to the design flexibility, structural integrity, versatile functionality, and resemblance to the native extracellular matrices (ECMs). However, existing reviews are limited in covering emerging trends and recent progress in ML and 3D electrospun scaffolds. A critical gap remains in synthesizing recent progress in 3D and ML electrospun scaffolds, which represent a transformative development in tissue engineering (TE). Therefore, this review specifically focused on the development of ML NFs scaffold and their application in TE and regenerative medicine (RM). The design of ML electrospun scaffolds allows the integration of 3D morphology, mechanical strength, biochemical signals, and antibacterial properties into a single structure. This review compiles recent advancements in electrospinning techniques, methodologies used for the fabrication of 3D scaffolds, and their applications in TE, including bone, cartilage, dental, and corneal tissue restoration, wound healing, and antibacterial activities. The literature reports that ML is becoming a key technology in RM.
Three-dimensional (3D) printed hydrogels have been recognized as a promising approach for cartilage tissue engineering (CTE) owing to their ability to accurately reproduce complex tissue architectures and to create a biomimetic microenvironment conducive to cell proliferation and differentiation. In this study, a novel 3D-printed composite scaffold composed of alginate, gelatin, and platelet-rich plasma (AG/PRP), incorporating magnesium-containing bioactive glass (Mg-BG), was developed (AG/PRP/BG). AG/PRP hydrogel formulations (10% w/v gelatin, 2% w/v alginate, and 0.2% w/v freeze-dried PRP) with varying Mg-BG concentrations (0.25, 0.5, and 1 wt%) were prepared to evaluate their effects on rheological behavior, printability, and biological performance. Rheological analyses revealed that all hydrogel formulations exhibited shear-thinning behavior, with the 0.5 wt% Mg-BG formulation (AG/PRP/BG2) demonstrating the highest print fidelity (printability index: 1.01 ± 0.02). Furthermore, bone marrow-derived mesenchymal stem cells (BM-MSCs) cultured on the 3D-printed scaffolds for 21 days showed enhanced chondrogenic differentiation in Mg-BG-containing group, as evidenced by increased SOX9 and collagen II gene expression, decreased collagen I gene expression, and elevated glycosaminoglycan (GAG) deposition. These findings demonstrate that the optimal incorporation of Mg-BG improves hydrogel viscosity, mechanical performance, and print fidelity, while promoting chondroinductive potential and maintaining high BM-MSCs viability, highlighting the suitability of AG/PRP/Mg-BG scaffolds as a robust candidate for advanced CTE applications.
Current bone tissue engineering scaffolds face challenges such as inadequate mechanical support, limited biocompatibility, and suboptimal osteogenesis for treating critical-sized bone defects. This study introduces a novel collagen/silk fibroin/carboxymethyl cellulose (CMC) scaffold integrated with ginsenoside Rg1-loaded Zeolitic Imidazolate Framework-8 (ZIF-8) nanoparticles (GINZIF-8), engineered to enhance the scaffold’s osteoinductive capacity and overall regenerative performance. Nanocomposite scaffolds were fabricated by incorporating ginsenoside Rg1–loaded ZIF-8nanoparticles into collagen/silk fibroin/ CMC matrices and processed via freeze-drying and glutaraldehyde crosslinking. Scaffold physicochemical properties, in vitro osteogenic responses of bone marrow–derived mesenchymal stem cells (BMMSCs), and in vivo bone regeneration in a rat critical-sized calvarial defect model were evaluated, with systemic safety analysis performed by liver histology and serum biochemical analyses. The constructs were fabricated to feature a fibrous, porous structure conducive to cell infiltration and nutrient diffusion. ZIF-8 nanoparticles were incorporated into the scaffold matrix, with successful encapsulation of ginsenoside Rg1 verified by Fourier Transform Infrared Spectroscopy (FTIR) analysis. The optimal formulation of the composite scaffolds (scaffolds loaded with 2 w/w% GINZIF-8) demonstrated improved mechanical properties (2.69 ± 0.14 MPa), and controlled ginsenoside Rg1 release (44.66 ± 7.84% over 7 days). In vitro assays confirmed that the GINZIF-8 nanoparticles significantly enhanced osteogenic differentiation of bone marrow mesenchymal stem cells (BMMSCs) potentially through activation of the Bone Morphogenetic Protein-2/SMAD signaling pathway. Additionally, the scaffolds exhibited antioxidant and anti-inflammatory effects, reducing oxidative stress and pro-inflammatory cytokine levels. In vivo studies revealed that the collagen/silk fibroin/CMC scaffolds loaded with 2 w/w% GINZIF-8 nanoparticles promoted substantial new bone formation (42.59 ± 3.70%) in a critical-sized defect model in rats, surpassing the performance of control scaffolds. Enhanced bone healing was attributed to the scaffold’s ability to support osteogenic differentiation, reduce oxidative stress, and modulate inflammation, with activation of the Phosphatidylinositol 3-Kinase/Protein Kinase B (PI3K/AKT) signaling pathway and inhibition of Glycogen Synthase Kinase-3 Beta (GSK3β) contributing to improved osteogenesis. These results demonstrate that Rg1-loaded ZIF-8 nanocomposite scaffolds effectively enhance BMMSC osteogenic differentiation and promote robust bone regeneration in vivo, while exhibiting excellent biocompatibility and systemic safety. Overall, the developed scaffolds represent a promising strategy for advanced bone tissue engineering applications.
The infrapatellar fat pad (IFP) is a rich source of mesenchymal stem cells (MSCs) with dual contributions from adipose and synovial tissues. The heterogeneity of IFP-derived MSCs and the lack of standardized isolation protocols, however, hinder consistent therapeutic outcomes. This study aimed to optimize collagenase-based isolation protocols for IFP-MSCs, with a focus on the effects of enzyme concentration and treatment duration on tissue digestion, cell origin, viability, and functional properties. IFP tissues harvested from patients undergoing knee arthroscopy were enzymatically digested using various collagenase concentrations (0.1-2%) and incubation times (2-48 h). Histological, immunohistochemical, flow cytometric, and functional assays were performed to evaluate tissue degradation, surface marker expression, colony-forming ability, and trilineage differentiation. Milder digestion conditions (2 h, 0.2-0.4% collagenase) preferentially extracted synovial membrane MSCs (CD55+ cells) and supported higher CFUs and chondrogenic/osteogenic differentiation. In contrast, prolonged digestion (48 h) led to increased cell yields and adipogenic differentiation, but reduced cell viability and percentage of synovial marker expression. In conclusion, enzymatic digestion parameters critically influence the cellular composition and regenerative potential of IFP-MSCs. Optimizing collagenase treatment conditions allows for a more selective, lineage-based MSC harvest, offering a practical strategy for tailored regenerative utilization of IFP-MSCs.
Wound healing is complex and still requires targeted therapies and advanced dressings despite current advances. Honey (HNY) is a multifunctional bioactive agent with demonstrated benefits across various phases of healing. Here, we present a wound stage-specific formulation strategy that combines a previously uncharacterized multiflora HNY from Monte Novo (Portalegre, Portugal) with gellan gum (GG) to develop gels for osmolarity-driven debridement and hydrogels for sustained release of bioactive components post-debridement, thereby enhancing therapeutic outcomes. The amount of sugars and phenolic compounds in the HNY were consistent with its hyperosmotic nature and anti-oxidant and anti-microbial activity. Gels were prepared using 0.1 and 0.2% GG with 50-90% HNY to reduce HNY's inherent flowability and enhance retention at the wound site, while maintaining extrudability. Hydrogels were formulated with higher GG (0.75 and 1.25%) to form mechanically stable crosslinked networks, and lower HNY (10-40%) to reduce osmolarity while preserving bioactivity. All gel formulations were extrudable and demonstrated effective water absorption. Those with higher GG and lower HNY exhibited increased viscosity, required greater force to flow, and displayed solid-like behavior, favoring wound retention. The 80% HNY-0.2% GG formulation balanced viscosity and osmotic strength, as confirmed by cytotoxicity results, while preserving anti-microbial activity indicating suitability for wound debridement. Hydrogels showed increased stiffness with higher GG and HNY while maintaining HNY's bioactive constituents. The 30% HNY-1.25% GG formulation demonstrated superior stability and sustained release of HNY components, including phenolic compounds at levels supporting anti-inflammatory and healing effects. Although initial HNY hydrogel exposure reduced cell metabolic activity, cells recovered over time, maintaining proliferation and migratory function. Collectively, these findings highlight the potential of GG/HNY gel/hydrogel formulations to support wound management across different healing stages, offering effective debridement through osmotic action and prolonged therapeutic support via controlled bioactive components release.
The prevalence of osteoporosis, a skeletal disorder characterized by reduced bone mass, microarchitectural deterioration, and increased fracture risk, poses a substantial global healthcare burden. Although animal models and two-dimensional cell cultures have been used to advance bone research, they do not completely replicate the multicellular interactions, extracellular matrix organization, and biomechanical environment of human bone, limiting their translational relevance. This review provides a critical synthesis of recent advances in bone organoid technology, emphasizing biological complexity, technical innovation, and relevance to osteoporosis modeling. Beyond summarizing progress, we distinguish validated capabilities from aspirational claims and identify the methodological gaps that must be addressed before bone organoids can reliably support drug screening, regenerative medicine, and precision approaches. Advances in stem cell biology, tissue engineering, and three-dimensional culture systems have enabled the use of self-organizing, multicellular organoids that reproduce key physiological and pathological features of bone. These systems model estrogen-deficiency-induced bone loss, glucocorticoid-associated osteoporosis, aging-related degeneration, and genetic susceptibility. By integrating osteogenic and endothelial components within biomimetic matrices, bone organoids can support mechanistic studies and pharmacological testing. However, their incomplete vascularization, limited mechanical fidelity, instability, and lack of standardized benchmarks restrict their translational readiness. Overcoming these barriers requires technological refinement, quantitative metrics, and regulatory alignment.
Dental pulp stem cells (DPSCs) represent an accessible and clinically relevant source of mesenchymal stem cells, and their derived exosomes have emerged as promising bioactive vesicles for cell-free applications in tissue engineering and regenerative medicine. However, reproducible and standardized methods for isolating and functionally validating DPSC-derived exosomes remain limited, particularly with respect to variability introduced during cell expansion across passages. Here, we describe a comprehensive, standardized, kit-based workflow for the isolation and purification of exosomes from DPSC-conditioned medium that is compatible with routine laboratory practice. This method enables consistent recovery of intact vesicles suitable for morphological, molecular, and functional characterization using commonly available techniques. Exosome identity is assessed based on morphology, size distribution, and marker expression, while biological activity is evaluated using a defined in vitro anti-inflammatory assay. To examine passage-related effects, exosomes derived from different DPSC passages are compared using this functional readout, providing a practical framework for assessing consistency during cell expansion. By integrating isolation, purification, characterization, and functional validation into a single workflow, this protocol offers a reproducible and scalable approach for generating functionally validated DPSC-derived exosomes for biomaterials research and regenerative applications.
Osteoarthritis (OA), the most prevalent joint disease and a leading cause of disability globally, has its disease burden inadequately captured by body mass index (BMI). As the sole quantified risk factor in current Global Burden of Disease estimates, BMI accounted for only 20% of OA burden. A critical limitation of BMI is its inability to distinguish fat distribution patterns, particularly abdominal adiposity, which is increasingly recognized as a key driver of metabolic and musculoskeletal pathologies. Herein, we hypothesize that anthropometric indicators reflecting central adiposity, such as average sagittal abdominal diameter (ASAD), may outperform BMI in predicting OA risk, especially when considering sex and age differences. This cross-sectional study analyzed 27,791 National Health and Nutrition Examination Survey participants (1999-2023) with complete OA diagnosis, anthropometric, and metabolic data. Participants were stratified by sex and age (40-year cutoff). Multivariable logistic regression, adjusted for confounders, estimated predictor-OA associations via standardized odds ratios (sORs), and these associations were evaluated by the area under the receiver operating characteristic curve (AUROC). Data were split into training (70%) and validation (30%) sets, with DeLong's test comparing different predictors against BMI. In the overall population, ASAD showed a stronger association with OA (sOR = 1.483) than BMI (sOR = 1.436), with comparable validation AUROC (ASAD: 0.857; BMI: 0.854). Sex-stratified analysis revealed that BMI was the optimal predictor for males (sOR = 1.466; validation AUROC = 0.844), while ASAD outperformed BMI in females (sOR = 1.486 vs. 1.450; validation AUROC = 0.865 vs. 0.863). Further age stratification revealed that in males under 40, both BMI (sOR = 1.261; validation AUROC = 0.750) and ASAD (sOR = 1.194; validation AUROC = 0.889) were the strongest predictors, and that ASAD (sOR = 1.490; validation AUROC = 0.769) and BMI (sOR = 1.482; validation AUROC = 0.736) remained strong for males aged 40 and above. In age-stratified analyses of females, ASAD showed the strongest consistent association with OA risk, both in participants under 40 (sOR = 1.472; validation AUROC = 0.801) and those aged 40 and above (sOR = 1.421; validation AUROC = 0.764). ASAD emerges as a superior predictor for females and a competitive population-level complement to BMI. BMI remains an optimal OA predictor for males. Within the National Health and Nutrition Examination Survey framework, these findings underscore the necessity of integrating abdominal adiposity metrics, particularly ASAD, into OA risk assessment to improve sex-specific prevention strategies.
Stress urinary incontinence (SUI) adversely impacts millions worldwide due to weakened pelvic floor muscles and urethral sphincter dysfunction. To date, there is a lack of effective non-surgical treatment for SUI, and no clear consensus has been reached on the optimal stem cell source under regenerative therapy. Existing studies have shown no precise molecular mechanisms underlying stem cell-mediated external urethral sphincter (EUS) regeneration. Therefore, we investigated the regenerative and reparative potential of our clinical-grade human amniotic fluid stem cells (hAFSCs) for treating SUI. We determined the immunophenotype, multi-differentiation potential, and secretome of AFSCs. Treated animals were grouped into sham, UI, phosphate buffer saline, and hAFSC groups. Pudendal nerve injury was created to induce SUI in female rats and treated with hAFSCs by administering them into the external urethral sphincter. Isolated AFSCs showed trilineage potential and expressed neuronal-specific markers such as Nestin, Tuj-1, MAP2, and GFAP. hAFSCs-treated group showed significantly (p < 0.01) improved leak point pressure, intercontractile interval, and total muscle cell proliferation numbers. hAFSCs showed elevated levels of VEGF, IL-8, TIMP-1, and TIMP-2. Histological assessment of bladder tissues reveals that hFASCS ameliorated lower ulceration and edema. Immunofluorescence staining and myogenic differentiation markers, i.e., Myf5, Myogenin, and MyoD, indicate the bladder tissue regenerating potential of hAFSCs. No hAFSC trafficking was observed in other tissues and organs. These findings highlight hAFSCs' potential as a novel therapy for SUI, warranting more extensive clinical trials to optimize dosing and long-term efficacy while addressing scalability and safety challenges in translating this regenerative approach to clinical practice.
Digital twin technology has emerged as a transformative innovation in healthcare, offering virtual replicas of physical entities at patient-level, equipment-level, and departmental-level that enable real-time monitoring, prediction, and optimisation. This narrative review synthesizes current evidence on digital twin maturity and clinical translation in radiology and radiotherapy. A comprehensive literature search was conducted across PubMed, Scopus, IEEE Xplore, and Web of Science databases for peer-reviewed articles published from 2018 onwards. The review reveals that digital twin applications in radiology remain predominantly experimental, with equipment-focused implementations (predictive maintenance, workflow optimization) showing greater maturity than patient-level applications. In radiology, emerging applications include personalised imaging protocol optimisation, predictive equipment maintenance, dose management, and workflow enhancement. In contrast, radiotherapy demonstrates more advanced patient-level digital twin integration, facilitating individualised treatment planning, real-time dose adaptation, treatment response prediction, and quality assurance. DT-aligned adaptive radiotherapy report improved local/locoregional control in the low-teens to ∼18% relative range, alongside clinically meaningful toxicity-risk reductions in selected endpoints, while maintaining lower radiation dose to organs. Key benefits include improved patient outcomes, reduced radiation exposure, enhanced treatment precision, and optimised resource utilisation. However, critical gaps persist in standardized validation frameworks, interoperability standards, and regulatory guidance. Implementation faces challenges including data integration complexity, computational requirements, regulatory uncertainties, and domain-specific barriers differing between radiology and radiotherapy contexts. Successful clinical translation requires addressing technical infrastructure gaps, establishing evidence-based validation protocols, and developing reimbursement mechanisms that recognize digital twin value. Digital twin technology demonstrates substantial potential for advancing precision medicine in imaging and radiation oncology.
Various factors such as infections, wounds, and comorbidities can disrupt the skin's physiological function. Moreover, skin lesions can result from radiation therapy. We aimed to create a new cosmetic formulation with pro-regenerative properties that is specifically designed for patients with sustained skin damage, such as those who have undergone radiation therapy. In the first stage, NE1 and IM2 peptides were synthesized, and hydrogels were prepared. IM2 is a derivative of the IM peptide, while NE1 contains the bioactive GHK sequence. In the next stage physicochemical analyses were conducted, including the evaluation of peptide stability and the developed composition's microbial purity and packaging compatibility. In addition, in vitro safety and activity assessments were implemented on human skin cells. In the final step, dermatological tests were conducted on the participants. The results indicate that the cosmetic composition is stable, possesses preservative properties, and is safe in both in vitro and in vivo studies. Peptide release studies show that within the first hours, approximately 75-80% of each peptide is released, ensuring a rapid onset of action. Analysis of cell migration indicate that both p407 and the designed composition stimulate migration of HaCaT keratinocytes in vitro. Dermatological tests did not show any irritant and sensitizing properties. Application analyses revealed that the designed composition effectively moisturizes and takes care of sensitive skin, alleviates redness and the effects of rough skin, eliminates the sensation of itching and the effects of skin tension, and soothes irritation. This formulation can be used for the daily care of sensitive, allergic, or irritated skin.
Systemic sclerosis (SSc) is characterized by progressive dermal fibrosis and microvascular dysfunction, and no approved therapy reliably reverses established skin fibrosis or durably restores microvascular perfusion. Adipose-derived stem cells (ASCs) possess anti-fibrotic, immunomodulatory, and vascular-related parameters properties, but their therapeutic impact in a strictly therapeutic (rather than preventive) SSc-like setting remains incompletely defined. Bleomycin-induced systemic sclerosis model was induced in male C57BL/6 mice by daily subcutaneous bleomycin injections (100 μg) into dorsal skin for 28 days. On day 14, mice received a single intralesional injection of ASCs (1 × 105 cells) or vehicle. At day 28, cutaneous perfusion was measured by laser Doppler perfusion imaging, and dorsal skin was analyzed by histology, hydroxyproline assay, RT-qPCR, and immunohistochemistry for CD34, α-SMA, and TNF-α. To support mechanistic interpretation, TGF-β1-stimulated dermal fibroblasts were co-cultured with ASCs and fibrosis-related gene expression was assessed. Intralesional ASC administration significantly attenuated bleomycin-induced dermal fibrosis, reducing dermal thickness (244.0-163.5 μm) and collagen area fraction (87.2-62.8%). Hydroxyproline content decreased from 0.187 to 0.121 μg/mg tissue. ASC treatment also suppressed profibrotic and inflammatory transcripts (α-SMA ~ 3.99-fold, TGF-β1 ~ 6.07-fold, TNF-α ~ 7.48-fold, IL-6 ~ 2.36-fold vs. BLM + PBS) and increased vascular responses transcripts (VEGF ~ 2.65-fold, CD34 ~ 1.28-fold vs. BLM + PBS). ASC co-culture suppressed profibrotic activation of TGF-β1-stimulated fibroblasts, reducing profibrotic expression (α-SMA ~ 2.5-fold, TGF-β1 ~ 3.5 -fold, and COL1A1 ~ 2.7-fold). A single intralesional ASC injection alleviated established bleomycin-induced dermal fibrosis and was associated with vascular-related changes in fibrotic tissue. These effects may involve paracrine-mediated suppression of TGF-β1-driven fibroblast activation, supporting ASCs as a promising regenerative strategy for systemic sclerosis skin disease.
Blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) is a rare congenital eyelid disorder that leads to drooping eyelids, narrowing of the palpebral fissures, and a characteristic facial appearance. It is a genetic condition, typically inherited in an autosomal dominant pattern, and is primarily caused by mutations in the FOXL2 gene, which is essential for eyelid development. Two types of BPES have been described: type II presents eyelid malformations and facial dysmorphism, while type I is additionally associated with premature ovarian insufficiency. If untreated, these features can lead to visual impairment and psychosocial consequences. A pronounced epicanthal fold may lead to a reduction in the visual field and contribute to visual disturbances, including strabismus. BPES is associated with underdeveloped or dysplastic levator palpebrae muscles, leading to four clinical features-telecanthus, ptosis, epicanthus inversus, and blepharophimosis-with reduced horizontal palpebral fissure length. A systematic review of the literature was performed to summarize current surgical management strategies for BPES. Surgical indications, outcomes, and complications were extracted and analyzed. Surgical treatment remains the standard of care for BPES. In recent years, numerous techniques have been developed to correct deformities including both classic and modern methods of epicanthoplasty, canthoplasty, and forehead muscle suspension using autologous and synthetic materials. Complication rates vary depending on technique and patient factors. This systematic review summarizes current surgical strategies, indications, outcomes and complications, highlighting modern approaches that optimize visual function and aesthetic outcomes. Surgical management of BPES is essential to restore eyelid function, improve the visual field, and achieve favorable cosmetic outcomes. A range of classical and contemporary techniques are available, and selection should be individualized. Modern approaches optimize both functional and aesthetic results, though further prospective studies are needed to standardize procedures and minimize complications.
Small extracellular vesicles (sEVs) derived from mesenchymal stem cells (MSCs) are emerging as potent acellular therapeutics; however, their rapid clearance hinders their clinical translation. To address this issue, 3D-bioprinted genipin-crosslinked gelatin (GECL) was engineered for human health. GECL hydrogels were functionalised with human umbilical cord MSC-derived sEVs (hUCMSC-sEVs) to create a bioactive wound-healing platform. These hydrogels demonstrated favourable physicochemical, mechanical, and biodegradable properties while providing an extracellular matrix (ECM)-mimetic environment conducive to tissue regeneration. MSCs were isolated from the umbilical cords, and their small extracellular vesicles (sEVs) were extracted and incorporated into gelatin-based hydrogels via 3D bioprinting. These sEV-loaded scaffolds were embedded in full-thickness wounds in mice, and healing was evaluated through macroscopic observation, histological analysis, collagen deposition, and angiogenesis assessment. Compared with the untreated controls, both the hydrogel-only (B) and sEV-loaded hydrogel (BE) groups significantly accelerated in vivo wound healing. Notably, the BE group achieved complete wound closure within 14 days, restoring the skin architecture, which closely resembled the native tissue with well-organised epidermal and dermal layers, optimal thickness, and skin appendages. Histological and ultrastructural assessments revealed an increased collagen type I deposition, a reduced α-smooth muscle actin (α-SMA) expression, and a robust neovascularisation. The TEM revealed tight junctions and active cellular infiltration, indicating scaffold integration and functional remodelling. Immunohistochemistry further revealed an upregulated CD31 expression with a balanced α-smooth muscle actin (α-SMA) expression, reflecting coordinated angiogenesis and myofibroblast regulation. These results highlight sEV-functionalised GECL hydrogels as robust and clinically translatable acellular therapeutic green products for accelerated wound closure and functional skin regeneration, advancing the fields of regenerative medicine and life expectancy.
The repair of bone defects remains a considerable challenge, primarily due to the lack of biomimetic hierarchical structures and the insufficient supply of bioenergy in implants. Inspired by the symbiotic structural relationship between mycelium and plants, we developed a biomimetic engineering strategy to construct mycelial bioceramics. This strategy enabled directing the growth of mycelium within bioceramic scaffolds, resulting in the spontaneous generation of a hierarchical structure. Such a hierarchical structure was attributed to the spontaneously microscale porous network of mycelium and the channel structure of the three-dimensional (3D) printed bioceramic scaffold. In addition, the mycelial bioceramics could release a variety of bioactive components, including glucose, calcium ions, and other ions. Hierarchical structure and bioactive components synergistically promoted cellular energy metabolism and osteogenic differentiation by enhancing glycolysis and the oxidative phosphorylation (OXPHOS) process. Furthermore, the mycelial bioceramics effectively activated the YAP/Piezo pathway, driving key mitochondrial biogenesis processes. The siYAP experiment combined with mRNA sequencing demonstrated that the elevated energy metabolism subsequently regulated osteogenic differentiation via PI3K-AKT signaling. In vivo studies using a rabbit femoral defect model demonstrated that mycelial bioceramics could improve cellular energy metabolism and ultimately enhance osteogenesis. In conclusion, the mycelial symbiotic strategy presents a novel approach in designing functional bioceramics for accelerating bone regeneration. Moreover, it may shed light on harnessing microorganisms for tissue engineering and regenerative medicine.