ObjectiveTo review the research progress of tissue engineered scaffolds and stromal-derived factor 1 (SDF-1) composite graft.
MethodsThe recent papers about SDF-1 with different kinds of tissue engineered scaffolds were reviewed and analyzed. The primary mechanism of SDF-1 homing function for stem cells was retrospected. The results of different kinds of tissue engineered scaffolds carrying SDF-1 for repairing the injured tissues and organs were reviewed.
ResultsIt is shown that SDF-1 combined with tissue engineered scaffolds will play a role of multipotent stem cells chemotaxis, however, the exact chemotaxis mechanism has not been fully understood. It still needs more researches of SDF-1 effects in vivo.
ConclusionAlthough some research progress has been made in regeneration in situ of tissue engineered scaffolds combined with SDF-1, it will need to further study on the mechanism of chemotactic functions of SDF-1 and its influence on proliferation and differentiation of cells.
Objective To find a kind of simple and effective method for purifying and label ing stromal vascular fraction cells (SVFs) so as to provide a theoretical basis for cl inical application of SVFs. Methods The subcutaneous adi pose tissue were harvested form volunteers. The adi pose tissue was digested with 0.065%, 0.125%, and 0.185% type I collagenase,respectively. SVFs were harvested after digestion and counted. After trypan blue staining, the rate of viable cells was observed. SVFs was labeled by 1, 1’-dioctadecyl-3, 3, 3’, 3’-2-tetramethy-lindocyanine perchlorate (DiI). The fluorescent label ing and growth was observed under an inverted fluorescence microscope. MTT assay was used to detect cell proliferation. Results The number of SVFs was (138.68 ± 11.64) × 104, (183.80 ± 10.16) × 104, and (293.07 ± 8.31) × 104 in 0.065% group, 0.125% group, and 0.185% group, respectively, showing significant differences among 3 groups (P lt; 0.01). The rates of viable cells were 91% ± 2%, 90% ± 2%, and 81% ± 2% in 0.065% group, 0.125% group, and 0.185% group, respectively, and it was significantly higher in 0.065% group and 0.125% group than in 0.185% group (P lt; 0.01), but no significant difference was found between 0.065% group and 0.125% group (P=0.881). Inverted fluorescence microscope showed that the cell membranes could be labeled by DiI with intact cell membrane, abundant cytoplasm, and good shape, but nucleus could not labeled. SVFs labeled by DiI could be cultured successfully and maintained a normal form. MTT assay showed that similar curves of the cell growth were observed before and after DiI labeled to SVFs. Conclusion The optimal collagenase concentration for purifying SVFs is 0.125%. DiI is a kind of ideal fluorescent dye for SVFs.
Objective To review research progress of adipose tissuederived stromal cells (ADSCs).Methods The recent articles on ADSCs were extensively reviewed, and the culture and differentiation ability of ADSCs were investigated.Results A population of stem cells could be isolated from adult adipose tissue, they were processed to obtain a fibroblast-like population of cells and could be maintained in vitro for extended periods with stable population doubling. The majority of the isolated cells were mesenchymal origin, with a few pericytes,endothelial cells and smooth muscle cells. ADSCs could be induced to differentiate intomultiple mesenchymal cell types, including osteogenic, chondrogenic, myogenic and adipogenic cells, they could also differentiate into nerve cells.Conclusion ADSCs can substitute mesenchymal stem cells and become an alternative stem cells source for tissue engineering.
The osteogenc potential of bone marrow has been proved by experiment. To investigate more in details, bone marrow was obtained from the trochanteric region of femur of NewZealand rabbit in 4 to 8 weeks old. After being cultured in vitro for one week, the hematopoietic component of the bone marrow had disappeared, thus the stromal cells were obtained. Then the stromal cells were subcultured in cultural fluid containing dexamethasone (10-8 mol/L) and natrium glycerophosphate (10mmol/L). Under the phasecontrast microscope, it was found that being cultured for 15 days. The stromal cells were lined up in one layer and late the secretion activity was increased and gradually transformed into multilayer structure and was congregated into diffused opaque clusters in twenty days. During culture, the cells were examined by tetracycline fluorescence label, histochemistry stains, transmission electron microscopy, scanning electron microscopy and energy dispersive X-ray microanalysis. The results showed that the morphological and biological characteristics of the cultured stromal cells derived from the bone marrow were similiar to those of osteoblasts and could synthesized mineralized new bone tissue in vitro.
ObjectiveTo evaluate the mechanism of stromal vascular fraction (SVF) promoting angiogenesis and tissue regeneration in tissue engineering chamber.
MethodsTwenty-four 6-month-old New Zealand white rabbits, male or female, weighing 2.5-2.8 kg, were selected. Thoracic dorsal arteriovenous bundle combined with collagen type I scaffold was transplanted to dorsal side, and wrapped by cylindrical hollow silicone chamber; all animals were randomly divided into the experimental group (n=12) and the control group (n=12). SVF was isolated from the back fat pads of rabbits in experimental group and labelled with DiI at 2 weeks after operation. The 1 mL cell suspension (1×106 cells/mL) and equal saline were injected into the chamber in experimental group and control group, respectively. The regenerative tissues were harvested for general observation and HE staining at 2 and 4 weeks after injection;and immunofluorescent staining was carried out in experimental group at 4 weeks.
ResultsAt 2 weeks after injection, the regenerative tissue was cylindrical; obvious vessel network and incompletely degradable collagen scaffold could be seen on the surface of the new tissue in 2 groups. The volume of new tissue was (0.87±0.11) mL in experimental group, and (0.72±0.08) mL in control group at 2 weeks, showing significant difference (t=2.701, P=0.011). At 4 weeks, little collagen scaffold could be seen on the surface in control group, but no collagen scaffold in experimental group; the volume of new tissue was (0.74±0.14) mL in experimental group, and (0.64±0.10) mL in control group, showing no significant difference (t=1.424, P=0.093). HE staining showed new mature vessels at 4 weeks, but no adipose tissue or fat lobulus formed in both groups; the capillary density was significantly higher in experimental group than in control group at 2 weeks (t=6.291, P=0.000) and at 4 weeks (t=5.445, P=0.000). The immunofluorescent staining found that SVF survived and located at the edge area after 4 weeks; the expressions of CD31 and DiI were positive in some endothelial cells.
ConclusionSVF can promote the angiogenesis and tissue regeneration in tissue engineering chamber, but it can not differentiate into adipocyte spontaneously without adipogenic microenvironment.
ObjectiveTo explore the methods of separation， culture， and identification of breast cancer stromal fibroblasts (BCSFs), which could build up a good basis for the further research of function. MethodsBreast cancer tissues were obtained during breast cancer operation， and were cut into pieces with size of 1 mm×1 mm×1 mm under aseptic conditions， then the pieces of the tissues were digested by collagenase Ⅰ and hyaluronidase. Finally the cells separated from the tissues incubated at 37 ℃ with 5% CO2 and 95% air humidified incubator. Morphological characteristics of the fibroblasts were observed under light microscope. The certain proteins were examined by immunohistochemistry （using CK， Vimentin， α-SMA, and TE-7 antibody） and flow cytometric analysis (CD34 and CD45). ResultsThe separated cells begin to attach to the wall of flask within 24 h and reached almost confluency in about 7 d to 10 d . According to identification, the successful rate of separation and culture of BCSFs was 90%（18/20）, and the characteristics of cells showed that morphological characteristics of the fibroblasts was flat spindle， rich cytoplasm, and a flat ovoid cystic nuclear. The fibroblasts in breast cancer tissues showed negative staining for cytokeratin， positive staining for vimentin， alpha-smooth muscle actin, and TE-7， and negative for CD34 and CD45 by flow cytometric analysis. ConclusionsThe fibroblasts in breast cancer tissues could be easily obtained by tissues cuting combined enzyme digestion and rocking technology in vitro. The present study provide an experimental foundation for further studies on fibroblasts in breast cancer.
Objective To investigate the expression of stromal cell-derived factor-1 (SDF-1) and its clinical significance in blood plasma of patients with breast tumor. Methods The level of SDF-1 protein was examined by enzyme linked immunosorbent assay (ELISA) in blood plasma of 26 patients with breast benign tumor and 52 patients with breast cancer. Results The SDF-1 protein in blood plasma was detected in both breast benign tumor patients and breast cancer ones. The level of SDF-1 protein in patients with breast cancer was higher than that in ones with breast benign tumor, and there was a statistical difference between them （P=0.000）. In patients with breast cancer, the level of SDF-1 protein in axillary lymph node (ALN) metastasis positive patients was significantly higher than that in ALN metastasis negative ones （P=0.036）. Conclusion The level of SDF-1 protein in blood plasma may be a specific tumor marker. Its level is correlated with lymph node involvement in breast cancer.
Objective To investigate the expression of stromal cell derived factor-1 ( SDF-1) and the effects of budesonide suspension for inhalation ( Pulmicort Respules) in mice with asthma. Methods Thirty Kunming female mice were randomly divided into three groups, ie. a control group, an asthma group, and a pulmicort treatment group. The asthma group and the pulmicort treatment group were sensitized with ovalbumin ( OVA) by a combination of intraperitoneal injection and repeated OVA intranasal challenges to establish mouse asthma model. The pulmicort treatment group received 100μL pulmicort by intranasal administration before OVA challenge. The immunohistochemistry was used to estimate the expression of SDF-1 in lung tissues. HE staining and Wright-Giemsa staining method were used to assess inflammatory infiltration in the airway and bronchoalveolar lavage fluid ( BALF) respectively. Results The expression of SDF-1 in the asthma group increased significantly compared with the control group ( 0.48 ±0.03 vs. 0.21 ± 0.02, Plt;0.05) , and significantly decreased after the intervention with pulmicort ( 0.29 ±0.01 vs. 0.48 ± 0.03, Plt; 0.05 ) . Compared with control group, the infiltration of inflammatory cells in airway was significantly enhanced in the asthma group, and attenuated in the pulmicort treatment group. The total number of inflammatory cells and eosinophil, lymphocyte, neutrophil counts in BALF increased significantly in the asthma group compared with the control group, and decreased significantly after pulmicort intervention. Conclusion SDF-1 may play an important role in the recruitment of inflammatory cells in asthmatic airway and pulmicort may relieve airway inflammation by decreasing the expression of SDF-1.
ObjectiveTo investigate the relationship between the level of stromal cell-derived factor-1 (SDF-1), internal carotid artery stiffness index, and non-arteritic anterior ischemic optic neuropathy (NAION) with macular edema (ME). MethodsA retrospective study. A total of 202 patients with NAION diagnosed by ophthalmic examination in Department of Ophthalmology, The Second Affiliated Hospital of Jiamusi University from January 2023 to January 2025 were included in the study. Based on the presence or absence of ME, the patients were divided into the NAION+ME group and the NAION group, with 94 and 108 cases respectively. A prediction model was constructed based on the influencing factors. To comprehensively evaluate the predictive value of SDF-1 level and carotid artery stiffness index for NAION with ME, a multidimensional analytical approach was employed. The diagnostic performance of individual and combined markers was assessed by constructing receiver operating characteristic (ROC) curves and calculating the area under the curve (AUC). Multivariate logistic regression analysis was performed to determine their independent predictive value. Stratified subgroup analyses were conducted to explore predictive differences across various populations. Cox proportional hazards regression models were established to evaluate long-term predictive value. Restricted cubic spline (RCS) analysis was applied to reveal potential nonlinear dose-response relationships. Mediation effect models were constructed to analyze the mediating role of carotid artery stiffness index in the association between SDF-1 level and NAION with ME. ResultsIn the NAION+ME group, systolic blood pressure (t=6.066), body mass index (t=2.804), disease duration (t=2.552), intraocular pressure (t=2.574), high-density lipoprotein (t=2.729), fasting blood glucose (t=2.022), glycosylated hemoglobin (t=7.235), SDF-1 level (t=14.319), and internal carotid artery stiffness index (t=2.633) were higher than those in the NAION group, while diastolic blood pressure was lower (P＜0.05). ROC curve analysis showed that the AUC of SDF-1 level combined with internal carotid artery stiffness index in predicting the risk of adverse prognosis was 0.894 [95% confidence interval (CI) 0.803-0.945], with a sensitivity of 87.98% and a specificity of 95.69%. Logistic regression analysis demonstrated significant independent correlations between SDF-1 level (OR=1.682, 95%CI 1.156-1.986), internal carotid artery stiffness index (OR=1.826, 95%CI 1.369-2.648), and the risk of ME in NAION patients (P＜0.05). Subgroup analysis revealed that elevated SDF-1 level and internal carotid artery stiffness index were associated with a higher risk of NAION with ME (Pfor trend＜0.05). RCS analysis demonstrated a nonlinear dose-response relationship between the continuous changes in SDF-1 level and internal carotid artery stiffness index and the risk of NAION with ME (P＜0.05). Mediation effect model analysis showed that internal carotid artery stiffness index played a mediating role between SDF-1 level and the risk of NAION with ME. ConclusionsSDF-1 level and internal carotid artery stiffness index are independent risk factors for ME in NAION patients. The combined detection of these two indicators holds significant value in predicting disease progression.
ObjectiveTo detect the content of stromal cell derived factor-1α(SDF-1α) in peripheral blood of patients with gastric adenocarcinoma (GC) and investigate its clinical significances.
MethodsThe contents of SDF-1αin the peripheral blood of 90 patients with GC were detected by enzyme-linked immunosorbent assay. The correlation of SDF-1αcontent with the clinicopathologic parameters and prognosis after operation were analyzed.
Results①The content of SDF-1αin the patients with GC[(6950.8±1131.3) ng/L] was significantly higher than that in the normal healthy volunteers[(5023.7±1103.8) ng/l, P=0.036].②The content of SDF-1αin the GC patients with distant metastasis[(8251.6±1042.5) ng/L] was significantly higher than that without distant metastasis[(6785.3±1025.0) ng/L, P < 0.001]. The contents of SDF-1αin the peripheral blood of patients with distant metastasis either in the liver (P=0.002) or in the lung (P=0.030) were significantly higher than those without distant metastasis (liver or lung).③The TNM stage was later (P < 0.001), lymph node metastasis was broader (P=0.018), invasion of tumor was deeper (P < 0.001), vascular invasion (P < 0.001) and lymphatic vessel invasion were present (P < 0.001), the contents of SDF-1αwere higer. Logistic regression analysis revealed that the depth of tumor invasion (OR=14.999, 95% CI 3.568-74.456, P=0.027) and distant metastasis (OR=0.186, 95% CI 0.610-2.014, P=0.026) were correlated with the high SDF-1αcontent.④The survival time of the patients with higher content of SDF-1αwas significantly shorter than that of the lower content of SDF-1α(P < 0.001). Cox proportial hazard regression model analysis demonstrated that TNM stage (RR=2.497, 95% CI 1.987-10.238, P=0.009), vascular invasion (RR=7.501, 95% CI 2.086-16.942, P=0.002), and high content of SDF-1α(RR=18.302, 95% CI 6.895-30.538, P=0.001) in the peripheral blood were the independent risk factors for survival of the patients with GC.
ConclusionHigh content of SDF-1αin peripheral blood might suggest the occurrence of lymph node metastasis, hepatic metastasis or lung metastasis and indicate the poorer prognosis of GC.
