Search

الملخص: أهداف البحث: تهدف الدراسة لتحديد محتوى البروتين الكلي، ونشاط مضادات الأكسدة، والقدرة الوقائية لمفرزات الخلايا الجذعية الوسيطة اللثوية البشرية في نموذج الإجهاد التأكسدي الناجم عن بيروكسيد الهيدروجين. طريقة البحث: تمت زراعة الخلايا الجذعية الوسيطة اللثوية البشرية للحصول على وسط مشروط (مفرزات)، ثم تم تجفيفها بالتجميد لإنتاج مفرزات مجففة بالتجميد. تم تحديد البروتين الكلي عن طريق فحص حمض البيكينكونيك والرحلان الكهربائي لهلام بولي أكريلاميد دوديسيل كبريتات الصوديوم لتحسين قياسات البروتين. تم قياس تركيز مضادات الأكسدة بواسطة اختبار ''أ بي تي اس''، في حين تم تحديد القدرة الوقائية للمفرزات المجففة بالتجميد ضد الإجهاد التأكسدي من خلال النشاط الأيضي لخلايا بانية العظم. تم تقسيم مجموعة الدراسة إلى مجموعة مراقبة (مستنبت) ومجموعة علاج المفرزات المجففة بالتجميد. النتائج: تحتوي المفرزات المجففة بالتجميد على تركيز بروتين يبلغ 2086.00 ± 0.20 ميكروغرام/مل، مع وزن جزيئي يبلغ 174، 74، 61، 55، و26 كيلو دالتون، والتي يعتقد أنها تسهل هجرة الخلايا، فضلا عن ربط السيتوكينات وعوامل النمو. كما قدمت المفرزات المجففة بالتجميد أعلى نشاط مضاد للأكسدة بنسبة 93.51% بتركيز 4.8 مجم/مل، مع قيمة تركيز مثبط50 تبلغ 2.08 مجم/مل. وقد ظهر أعلى نشاط استقلابي للخلية في مجموعة العلاج بالمفرزات المجففة بالتجميد 1.25 ملغم/مل. جميع تركيزات المفرزات المجففة بالتجميد للخلايا الجذعية الوسيطة اللثوية البشرية - تخفف من التأثير الضار للإجهاد التأكسدي الناجم عن بيروكسيد الهيدروجين. الاستنتاجات: يمكن أن تحافظ المفرزات المجففة بالتجميد التي تم الحصول عليها من الخلايا الجذعية الوسيطة اللثوية البشرية على النشاط الأيضي في خلايا بانيات العظم كحماية ضد إجهاد بيروكسيد الهيدروجين. Abstract: Objective: To determine total protein content, antioxidant activity, and protective ability of lyophilized human gingival mesenchymal stem cells (hGMSCs)-secretome in hydrogen peroxide (H2O2) induced oxidative stress model. Methods: Human GMSCs were cultured to obtain a conditioned medium (secretome), then lyophilized to produce lyosecretome. Total protein was determined by bicinchoninic acid assay (BCA) and SDS-PAGE to improve protein measurements. Antioxidant concentration was measured by ABTS assay, while the protective ability of lyosecretome against oxidative stress was determined by the metabolic activity of osteoblast cells. The study group was divided into a control group (culture medium) and a lyosecretome treatment group (0.0; 0.157, 0.313, 0.625, 1.25, 2.5, 5, and 10 mg/mL + H2O2). Results: Lyosecretome had a protein concentration of 2086.00 ± 0.20 μg/ml, with a molecular weight of 174, 74, 61, 55, and 26 kDa, which are thought to facilitate cell migration, as well as bind cytokines and growth factors. Lyosecretome also provided the highest antioxidant activity of 93.51% at a concentration of 4.8 mg/ml, with an IC50 value of 2.08 mg/ml. The highest cell metabolic activity (79.53 ± 2.41%) was shown in the 1.25 mg/ml lyosecretome treatment group. All concentrations of hGMSC-lyosecretome attenuate the adverse effect of H2O2-induced oxidative stress. Conclusion: Lyosecretome obtained from hGMSCs can maintain metabolic activity in osteoblast cells as protection against H2O2 oxidative stress.

Copyright of Journal of Taibah University Medical Sciences is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Recently, 3D-printed scaffolds for the controlled release of mesenchymal stem cell (MSC) freeze-dried secretome (Lyosecretome) have been proposed to enhance scaffold osteoinduction and osteoconduction; coprinting of poly(ε-caprolactone) (PCL) with alginate hydrogels allows adequate mechanical strength to be combined with the modulable kinetics of the active principle release. This study represents the feasibility study for the sterile production of coprinted scaffolds and the proof of concept for their in vitro biological efficacy. Sterile scaffolds were obtained, and Lyosecretome enhanced their colonization by MSCs, sustaining differentiation towards the bone line in an osteogenic medium. Indeed, after 14 days, the amount of mineralized matrix detected by alizarin red was significantly higher for the Lyosecretome scaffolds. The amount of osteocalcin, a specific bone matrix protein, was significantly higher at all the times considered (14 and 28 days) for the Lyosecretome scaffolds. Confocal microscopy further confirmed such results, demonstrating improved osteogenesis with the Lyosecretome scaffolds after 14 and 28 days. Overall, these results prove the role of MSC secretome, coprinted in PCL/alginate scaffolds, in inducing bone regeneration; sterile scaffolds containing MSC secretome are now available for in vivo pre-clinical tests of bone regeneration. [ABSTRACT FROM AUTHOR]

Recently, 3D-printed scaffolds for the controlled release of mesenchymal stem cell (MSC) freeze-dried secretome (Lyosecretome) have been proposed to enhance scaffold osteoinduction and osteoconduction; coprinting of poly(ε-caprolactone) (PCL) with alginate hydrogels allows adequate mechanical strength to be combined with the modulable kinetics of the active principle release. This study represents the feasibility study for the sterile production of coprinted scaffolds and the proof of concept for their in vitro biological efficacy. Sterile scaffolds were obtained, and Lyosecretome enhanced their colonization by MSCs, sustaining differentiation towards the bone line in an osteogenic medium. Indeed, after 14 days, the amount of mineralized matrix detected by alizarin red was significantly higher for the Lyosecretome scaffolds. The amount of osteocalcin, a specific bone matrix protein, was significantly higher at all the times considered (14 and 28 days) for the Lyosecretome scaffolds. Confocal microscopy further confirmed such results, demonstrating improved osteogenesis with the Lyosecretome scaffolds after 14 and 28 days. Overall, these results prove the role of MSC secretome, coprinted in PCL/alginate scaffolds, in inducing bone regeneration; sterile scaffolds containing MSC secretome are now available for in vivo pre-clinical tests of bone regeneration.