Adipose-drive stem cells (ASCs) are similar to Mesenchymal stem cells (MSCs), which are considered as multipotent progenitors that have capable of immunomodulation, self-renewal and differentiation into multiple cell lineages that can be used in cancer therapy or immune diseases. Dimethyl fumarate (DMF) is one of the important drugs with modulating function for treatment of various immune disease. The primary aim of this study was to investigate how dimethyl fumarate influences the immunomodulatory profile and functional properties of adipose‑derived mesenchymal stem cells under inflammatory conditions. Adipose-stimulating stem cells (ASCs) were isolated from the abdominal adipose tissue of ten patients via liposuction, and DMF was added after inducing inflammatory conditions with inflammatory cytokines such as IFN-γ and TNF-α. Cell culture and RT-PCR were performed to measure the expression of immunomodulatory factors such as Galectin-1/3, HGF, HO-1, CXCL-8, and IL-6. Flow cytometry for mesenchymal stem cell nature and confirmation was done. Fat and bone staining was performed to test the functional differentiation of these cells. ASCs treated with DMF at 0.01-100 μM concentration at 2 days showed the enhancement of CXCL-8 and IL-6 gene expression notably, whereas, HGF, and Galectin-1 reduced. Moreover, Galectin-3 and HO-1 depicted no significant difference compared to the control group. Further, Alizarin Red and Oil Red staining verified the change of MSCs toward adipose and osteogenic differentiation. ASCs are positive for markers including CD73+, CD90+, and CD105+ antigens by abs staining conjugated to fluorescence dye. Our study confirmed that ASCs combined with DMF have increasingly important roles through immune-modulatory properties by secreting mediators, which can suppress, improve, alter, or change the microenvironment of disease. Despite ongoing progress, the underlying stimulatory and regulatory mechanisms remain incompletely understood. The gap between pre‑clinical findings and clinical applications highlights the need for further investigation. Extensive research is still required to elucidate how therapeutic interventions achieve efficacy, ensure safety, and identify optimal strategies for targeting different organs in combination with other pharmacological agents.

Allogenic bone grafting and bone banking have become vital components of modern orthopaedic reconstruction, addressing bone loss following trauma, infection, tumor resection, and revision arthroplasty. Despite expanding clinical use, a comprehensive overview of global research productivity and collaboration in this domain has not been previously undertaken. A bibliometric analysis was conducted using the Scopus database (2000-2025) with defined search terms related to bone banking and allogenic bone grafting in orthopaedics. Data were analyzed using Scopus analytics and VOSviewer for publication trends, source impact, geographic distribution, authorship, funding patterns, and keyword co-occurrence networks. A total of 3497 documents were identified, showing steady publication growth. The majority were original research articles (79.2%). The United States led in publication output (37.7%), followed by China and Italy. Clinical Orthopaedics and Related Research and Spine were the most productive journals. Keyword mapping revealed core themes in revision arthroplasty, spinal fusion, limb reconstruction, and bone defect management. Global research on allogenic bone and bone banking demonstrates robust growth, interdisciplinary collaboration, and emerging regional contributions. Future efforts should emphasize standardization, outcome-based studies, and integration of biomaterials and regenerative technologies to enhance the safety and sustainability of bone banking worldwide.

Previous studies have suggested that H-Y antigens may influence immunological outcomes in corneal transplantation. This study analyses the influence of donor-recipient gender mismatch in DMEK on postoperative outcomes. This single-center retrospective study included 2521 eyes (1712 patients) that underwent DMEK between January 2012 and December 2021. Patients were divided into four groups according to donor-to-recipient gender combinations (male-to-female, male-to-male, female-to-male, female-to-female) and were compared in terms of graft failure rate, visual acuity (VA), central corneal thickness (CCT), and endothelial cell count (ECC) preoperatively and at 1, 12 and 36months postoperatively, as well as in terms of postoperative complications. Of the 2521 cases, 1647 (65%) were sex-mismatched (male-to-female or female-to-male) and 874 (35%) were sex-matched (female-to-female or male-to-male). Intraoperative preparation time of the grafts (p = 0.78) and difficulties in the intraoperative handling (p = 0.43) were comparable in all groups. Re-bubbling or re-keratoplasty was required with comparable frequency (p = 0.84; p = 0.61 respectively). The occurence of primary or secondary graft failure was not statistically significant different between the groups (p = 0.57). No confirmed case of immunological graft rejection occurred. Functional postoperative outcomes for ECC (p = 0.18 after 1month, p = 0.13 after 12months, p = 0.27 after 36months), CCT (p = 0.58 after 4-6weeks, p = 0.82 after 12months, p = 0.78 after 36months) and VA (p = 0.64 after 4-6weeks, p = 0.47 after 12months, p = 0.39 after 36months) were comparable in all groups at the follow-up. In this retrospective cohort, no documented immune graft rejection was observed, and sex mismatch did not influence intraoperative handling or postoperative interventions. However, prospective studies with standardized immunologic assessment are needed to clarify the role of donor-recipient sex mismatch in DMEK outcomes.

Bone regeneration remains a major clinical challenge due to the limited healing capacity of large bone defects and the limitations of conventional grafting or cell-based therapies. Exosomes, nanosized extracellular vesicles secreted by diverse cell types, have emerged as promising cell-free mediators of osteogenesis, angiogenesis, and immune regulation. However, the therapeutic efficacy of native exosomes is constrained by low yield, rapid clearance, and limited targeting. Because effective bone regeneration is inherently multi-factorial-requiring biomechanical stability, vascularization, and an instructive ECM and cellular microenvironment-engineered exosomes should be regarded as enabling components within integrated regenerative systems rather than a standalone solution. Recent advances in engineered exosomes (EExos) have opened new frontiers in bone tissue regeneration by enabling precise design, biofunctionalization, and targeted delivery. Engineering strategies-ranging from genetic modification of donor cells to chemical conjugation, hybrid nanocarrier formation, and controlled cargo loading-have been employed to enhance the osteoinductive and osteoconductive potential of exosomes. Furthermore, incorporation of EExos into smart delivery systems, such as hydrogel scaffolds, 3D-printed matrices, and bone-targeting ligands, offers sustained release and localized therapeutic effects within the bone microenvironment. This review comprehensively summarizes the latest developments in the design, delivery, and functional optimization of EExos for bone regeneration. Mechanistic insights into their roles in promoting bone remodeling, angiogenesis, and immune modulation are discussed alongside current translational progress, manufacturing challenges, and regulatory considerations. Finally, emerging directions-such as AI-assisted exosome engineering, CRISPR-based programming, and bioprinting-integrated therapies-are highlighted as transformative pathways toward personalized and clinically translatable bone regenerative medicine.

Descemet's membrane is the specialized extracellular matrix located between corneal stroma and endothelium. This basement membrane provides the biomechanical cues that sustain endothelium viability and function, making it an optimal cell scaffold. The present work describes a new decellularization protocol to remove the cellular components and obtain an acellular scaffold from Descemet's membrane. To induce cell lysis and eliminate all cytoplasmic and nuclear material, Descemet's membranes isolated from donor corneas were subjected to osmotic (hypotonic) shock. The efficiency of the decellularization process was evaluated by the quantification of total residual DNA and analysis by gel electrophoresis. Nuclei removal and extracellular matrix integrity after treatment were verified by histological analysis. In particular, the maintenance of collagen, glycoproteins, perlecan and elastin was analyzed in decellularized tissues compared to untreated controls. DNA quantification showed a 99% reduction of total DNA amount in decellularized Descemet's membranes compared to control ones, with only 28.9 ± 9.86ng DNA/mg dry tissue residual. Furthermore, the agarose gel electrophoresis and the absence of visible nuclei after decellularization confirmed the efficiency of the process. Histological analyses showed that the composition of the extracellular matrix was not modified by the process. The decellularization protocol is effective in obtaining a Descemet's membrane that is depleted of donor DNA. Furthermore, the treatment preserves tissue matrix components. Descemet's membrane is already prestripped and provided by eye banks; therefore, a decellularized Descemet's membrane represents a valid candidate as a safe scaffold for intraocular surgery as, for example, in the treatment of refractory macular holes.

To evaluate the treatment success and reversal rate of corneal allograft rejections in post-penetrating keratoplasty patients. Retrospective cohort study was performed in post-penetrating keratoplasty patients treated for corneal allograft rejection between September 2015 and August 2024 in a tertiary eye center in Debrecen, Hungary. Indication of keratoplasty, onset of the allograft rejection, best corrected visual acuity (BCVA), corneal transparency on slit lamp examination and applied treatment were recorded. In our institution's patient registry, 68 allograft rejection episodes were found. Before the rejection, all grafts were completely transparent. The rejection episodes occurred median 21months after surgery. Diagnosis and treatment took place median 5days after the onset of symptoms. Allograft rejection could be successfully reversed in 48 of the total 68 cases (70.6%). Treatment was adjusted individually and all patients received local treatment; 16 received only local and 52 received combined systemic and local corticosteroids resulting in a similar rejection reversal rate (13/16 vs. 35/52; p = 0.359). Comparing first (47) and repeat grafts (21), there were no significant differences between the treatment success rate (33/47 vs. 15/21; P = 1.000) and the frequency of combined local-plus-systemic treatment (34/47 vs. 18/21; p = 0.355). Before the rejection episodes, BCVA was 0.40 ± 0.30, which decreased following the rejection (0.28 ± 0.28; p < 0.001). This reduction in BCVA was observed in successfully treated cases as well (0.44 ± 0.28 vs. 0.38 ± 0.27; p = 0.008). Our dexamethasone-based local treatment demonstrated similar effectiveness in reversing corneal allograft rejections in patients underwent penetrating keratoplasty compared to data on prednisolone by other studies. However, systemic steroid augmentation might be needed more frequently when using topical dexamethasone.

Bone morphogenetic protein (BMP) stimulated osteoinduction is critical for bone regeneration. Human demineralized bone matrix (DBM) has been one of the most widely used bone graft substitutes, but its osteoinductive potential is weak and clinical effectiveness limited in part due to ineffective processing methods. Here, we describe a novel process designed to enhance allograft bioactivity by increasing the bioavailability of the native BMPs present within the matrix the product of which we call Natural Matrix Protein® (NMP®). BMP-7 release was quantified from NMP and DBM prepared from both bovine and human cortical bone. In both species, NMP significantly increased BMP-7 levels in acidic and physiologic extracts compared to DBM. NMP also demonstrated sustained release of BMP-7 and total protein for up to 12 weeks in simulated body fluid. Osteoinductive potential was evaluated in vitro using C2C12 cells and osteoinductivity in vivo in the athymic rat model. Direct treatment of cells with NMP in vitro produced a greater than tenfold increase in alkaline phosphatase activity at 40 mg/mL. In vivo, human NMP showed increased osteoinductivity compared to human DBM histologically, and the recovered NMP explants had significantly more mineralized bone and a higher bone volume fraction compared to DBM and to Infuse® Bone Graft (105µg rhBMP-2 on an absorbable collagen sponge) as measured by microCT. These findings demonstrate that the novel NMP process reproducibly increases BMP-7 bioavailability, that NMP implants produce sustained BMP and protein release and have marked increase in osteoinductive activity.

Regeneration of articular cartilage disorders is one of the critical challenges in musculoskeletal medicine. Tissue engineering could represent a therapeutic option to support cartilage regeneration. Natural and biological materials are appropriate for fabricating tissue engineering scaffolds because of their similarity to natural tissues. The properties of amniotic membranes, including low immunogenicity, anti-inflammatory role, cell loading capability, expression of various growth factors, and chondroprotective effect, make them an interesting option for cartilage regeneration. This review studied the structure of articular cartilage and potential applications of the human amniotic membrane (AM) for articular cartilage regeneration. In addition, processing and decellularization methods of AM and the most common forms of amniotic membrane used in cartilage regeneration, including sheet, injectable form, and 3D forms, were studied. This review highlights the benefits of amniotic membrane applications in cartilage regeneration and clinical trial studies.

This study aimed to determine the most feasible perinatal tissue for Good Manufacturing Practice (GMP)-compliant banking of mesenchymal stromal-like cells (MSC-like cells). It was hypothesized that amniotic fluid collected during cesarean section would yield lower contamination rates and greater processing feasibility compared with other perinatal tissues. This prospective observational study was conducted at a tertiary university hospital and included 32 healthy term pregnancies. A total of 160 perinatal samples-amniotic fluid, amniotic membrane, umbilical cord, intact placenta, and placental fragments-were obtained. A validated feasibility scoring system evaluated material acquisition difficulty, transportation logistics, storage duration, and processing complexity. Samples were stratified by delivery mode (cesarean section vs. vaginal delivery) and collection timing (within vs. outside laboratory working hours). Stem cell isolation, sterility assessment, and immunophenotypic characterization were performed. Due to the absence of trilineage differentiation assays and maternal-fetal origin confirmation, the isolated cells were defined as MSC-like cells rather than definitive fetal MSCs. Statistical analyses were performed using chi-square and Mann-Whitney U tests (p < 0.05). Samples collected via cesarean section demonstrated significantly lower rates of blood contamination (25.8% vs. 60.0%, p < 0.001) and bacterial contamination (25.8% vs. 60.0%, p < 0.001) compared with those from vaginal deliveries. Amniotic fluid achieved the highest acquisition score, required no enzymatic digestion, and had the shortest median isolation time (45min). It exhibited the lowest overall contamination and was the most suitable source for GMP-oriented MSC-like cell processing. Conversely, intact placenta and placental fragments showed the highest contamination rates, longest enzymatic processing times, and greatest logistical burden. While collection timing affected storage duration and workflow continuity, tissue type and delivery mode were the dominant determinants of feasibility. Cesarean section-derived amniotic fluid appears to be the most practical, sterile, and processing-efficient perinatal source for GMP-adapted MSC-like cell banking within the evaluated parameters. These results support its prioritization in the development of standardized collection and processing protocols for perinatal stromal cell applications in regenerative medicine.

The current investigation aimed to exhibit the impact of lyophilization media of frozen spermatozoa on the fertility potential of buffalo spermatozoa as indicated by comet assay and ICSI. Semen specimens were centrifugated at 700 × g for 20min using percoll gradient (45-90%), double washed in Tyrode's albumen lactate pyruvate (TALP) and diluted in the lyophilization media (media 1, 2, 3 and 4), correspondingly. Cooling of the diluted sperm cells in vapor of liquid nitrogen. Frozen samples were instantly put into the lyophilizer (-55°C, pressure 0.001 Mbar). After 24h of lyophilization, the semen specimens were kept for three months at 4°C. Frozen-dried semen was re-hydrated at room temperature in of milli-Q water(100 µL). Comet assay results of the frozen-dried semen exhibited that the TCM medium exhibited the lowest % of DNA deterioration [6.17] and the superior % of embryonic developmental rate,while Tris-EDTA medium exhibited the highest % of DNA deterioration [13.09]. The lowest successful % of ICSI exhibited upon using EGTA and EDTA media. It could be concluded that ICSI of frozen-dried spermatozoa upon using TCM medium provides the highest % of embryonic expansion. Also, Tris-EGTA and Tris-EDTA media exhibited the lowest successful percent.