Metabolic signatures of tear extracellular vesicles caused by herpes simplex keratitis.

PURPOSE: Herpes simplex keratitis (HSK), caused by type 1 herpes simplex virus (HSV) reactivation, is a severe infectious disease that leads to vision loss. HSV can trigger metabolic reprogramming in the host cell and change the extracellular vesicles (EV) cargos; however, little is known about the EV metabolic signatures during ocular HSV infection. Here, we aimed to depict the EV-associated metabolic landscape in HSK patients' tears. METHODS: We collected 82 samples from 41 participants with unilateral HSK (contralateral unaffected tears were set as negative control), including subtype cohorts of 13 epithelial, 20 stromal, and 8 endothelial HSK. We isolated tear EVs via our previously established platform and conducted metabolic analysis using LC-MS/MS. The metabolic signatures for recognizing HSK and subtypes were assessed through differential analysis and machine learning algorithms. RESULTS: Hypopsia and increased extracellular CD63 levels were observed in affected eyes. We identified 339 metabolites based on sEVs isolated from tears. Differential analysis revealed alterations in energy and amino acid metabolism, as well as the infectious microenvironment. Furthermore, we observed dysregulated metabolite such as methyldopa, which is associated with inappropriate neovascularization and corneal sensation loss, contributing to the HSK severity particularly in the stromal subtype. Moreover, machine learning classification also suggested a set of EV metabolic signatures that have potential for pan-keratitis detection. CONCLUSIONS: Our findings demonstrate that tear EV metabolites can serve as valuable indicators for comprehending the underlying pathological mechanisms. This knowledge is expected to facilitate the development of liquid biopsy means and therapeutic target discovery.

[Biomechanical study of polymethyl methacrylate bone cement and allogeneic bone for strengthening sheep vertebrae].

OBJECTIVE: To investigate the feasibility and mechanical properties of polymethyl methacrylate (PMMA) bone cement and allogeneic bone mixture to strengthen sheep vertebrae with osteoporotic compression fracture. METHODS: A total of 75 lumbar vertebrae (L 1-L 5) of adult goats was harvested to prepare the osteoporotic vertebral body model by decalcification. The volume of vertebral body and the weight and bone density before and after decalcification were measured. And the failure strength, failure displacement, and stiffness were tested by using a mechanical tester. Then the vertebral compression fracture models were prepared and divided into 3 groups ( n=25). The vertebral bodies were injected with allogeneic bone in group A, PMMA bone cement in group B, and mixture of allogeneic bone and PMMA bone cement in a ratio of 1∶1 in group C. After CT observation of the implant distribution in the vertebral body, the failure strength, failure displacement, and stiffness of the vertebral body were measured again. RESULTS: There was no significant difference in weight, bone density, and volume of vertebral bodies before decalcification between groups ( P>0.05). After decalcification, there was no significant difference in bone density, decreasing rate, and weight between groups ( P>0.05). There were significant differences in vertebral body weight and bone mineral density between pre- and post-decalcification in 3 groups ( P<0.05). CT showed that the implants in each group were evenly distributed in the vertebral body with no leakage. Before fracture, the differences in vertebral body failure strength, failure displacement, and stiffness between groups were not significant ( P>0.05). After augmentation, the failure displacement of group A was significantly greater than that of groups B and C, and the failure strength and stiffness were less than those of groups B and C, the failure displacement of group C was greater than that of group B, and the failure strength and stiffness were less than those of group B, the differences between groups were significant ( P<0.05). Except for the failure strength of group A ( P>0.05), the differences in the failure strength, failure displacement, and stiffness before fracture and after augmentation in the other groups were significant ( P<0.05). CONCLUSION: The mixture of allogeneic bone and PMMA bone cement in a ratio of 1∶1 can improve the strength of the vertebral body of sheep osteoporotic compression fractures and restore the initial stiffness of the vertebral body. It has good mechanical properties and can be used as one of the filling materials in percutaneous vertebroplasty.

A bone substitute composed of polymethyl-methacrylate bone cement and Bio-Gene allogeneic bone promotes osteoblast viability, adhesion and differentiation.

BACKGROUND: Increasing reports on new cement formulations that address the shortcomings of PMMA bone cements and various active components have been introduced to improve the biological activity of PMMA cement. OBJECTIVE: Evaluating the biological properties of PMMA cements reinforced with Bio-Gene allogeneic bone. METHODS: The MC3T3-E1 mouse osteoblast-like cells were utilized to determine the effects of Bio-Gene + PMMA on osteoblast viability, adhesion and differentiation. RESULTS: The combination of allogeneic bone and PMMA increased the number of adherent live cells compared to both control group and PMMA or Bio-Gene group. Scanning electron microscopy observed that the number of cells adhered to Bio-Gene + PMMA was larger than Bio-Gene and PMMA group. Compared with the control and PMMA or Bio-Gene group, the level of ALP and the number of calcium nodules after osteoinduction was remarkably enhanced in Bio-Gene + PMMA group. Additionally, the combination of Bio-Gene and PMMA induced the protein expression of osteocalcin, osterix and collagen I. CONCLUSION: The composition of PMMA and allogeneic bone could provide a more beneficial microenvironment for osteoblast proliferation, adhesion and differentiation. PMMA bone cement reinforced with Bio-Gene allogeneic bone may act as a novel bone substitute to improve the biological activity of PMMA cement.