Objective To establish a method of isolating and culturing adult human bloodderived mesenchymal stem cells(MSCs) and to investigate their osteogenic potential in vitro. Methods Thirty peripheral blood sampleswere collected from 30adult volunteers(15 ml per person).Adult human MSCs derived from peripheral blood were isolated from the lymphocyte separation fluid fraction of mononuclear cells, cultured in α-Modified Eagle’s Medium with low glucose containing 20% fetal bovine serum, and proliferated through a process of subculturing. The phenotype of MSCs was analyzed with flow cytometry. For in vitro osteogenic differentiation, MSCs from the second passage grew in the presence of osteogenic supplements (100 nmol/L dexamethasone,10 mmol/L β-glycerophosphate,50 μmol/L vitamin C, and 10 nmol/L 1,25-2-hydroxide vitamin D3). In the fifth passage cells, the activity of alkaline phosphatase, the expression level of collagen typeI, osteocalcin and osteonectin were determined. And the calcium tubercle formation would be examined after the continual one-month culture of the fifth passage. Results MSCs exsited in the pheripheral blood of adult human. And the clone forming efficiency of blood-derived MSCs was 0.27±0.22/106 mononuclear cells. The MSCs expressed CD44,CD54,CD105,and CD166，but did not CD14, CD34, CD45,and CD31.Under the function of osteogenic supplements, the MSCs were found to be higher activity of alkaline phosphatase and higher expression levels of collagen type Ⅰ, osteocalcin and osteonectin. And the calcium tubercle formation was examined throughtetracycline fluorescence labeling method. Conclusion The isolation and cultureconditions established for adult human MSCs may select a distinct population of peripheral blood-derived adherent cells. Adult human blood-derived MSCs possess osteogenic potential in vitro, and may be used as seed cells for bone tissue engineering.
ObjectivesTo explore the changes of some peripheral blood cells related to inflammation in patients with non-arteritis central retinal artery occlusion (NA-CRAO). MethodsA retrospective clinical study. From July 2019 to July 2021, a total of 218 patients with NA-CRAO hospitalized (NA-CRAO group) in Department of Ophthalmology, Xi'an People's Hospital (Xi'an Fourth Hospital) and 218 patients with routine physical examination (control group) during the same period were included in the study. There were no significant differences in age (t=0.60), sex composition ratio (χ2=0.83) and body mass index (t=0.77) between the two groups (P＞0.05). 0.2 ml fasting peripheral blood was collected from the subject, and white blood cells (WBC), neutrophils (NEUT), lymphocytes (LYMPH), red blood cells (RBC), RBC distribution width (RDW), platelets (PLT), mean PLT volume (MPV), and large PLT ratio (PLCR) were detected. The NEUT/LYMPH ratio (NLR) and PLT/LYMPH ratio (PLR) were calculated. t test was used to compare measurement data between groups. Multiple logistic regression analysis was performed for blood cells with P＜0.05. The receiver operating characteristic curve (ROC curve) was used to calculate the area under the curve (AUC) and 95% confidence interval (95%CI) of each inflammatory indicator, and the optimal cutoff value was determined according to the Jorden index (sensitivity+specificity－1). ResultsCompared with control group, WBC, NEUT, NLR, RDW, PLR were increased in NA-CRAO group, while RBC and LYMPH were decreased, with statistical significance (t=9.68, 12.43, 9.47, 3.64, 5.54, 5.18, 0.46; P＜0.001). There was no significant difference in PLT, MPV and PLCR between the two groups (t=0.32, 1.56, 0.84; P＞0.05). Multivariate logistic regression analysis showed that NLR was a possible risk factor for the occurrence of NA-CRAO (odds ratio=2.51, 95%CI 0.780-0.859, P=0.031). ROC curve analysis showed that the AUC predicted by NLR was 0.819, the optimal critical value was 3.05, and the sensitivity and specificity were 59.2% and 92.7%, respectively. ConclusionsIn peripheral blood cells of NA-CRAO patients, NEUT is significantly increased and LYMPH is decreased. NLR is a possible risk factor for NA-CRAO.