Chondrogenic differentiation of bone marrow‑derived stem cells cultured in the supernatant of elastic cartilage cells.
Zhang Xiaodie,Xue Ke,Zhou Jia,Xu Peng,Huang Huizhen,Liu Kai
Molecular medicine reports
Repair of cartilage defects remains a challenge for surgeons, owing to its poor self‑repairing capacity. Cartilage tissue engineering, particularly marrow stem cell‑based cartilage regeneration, provides a promising option for the regeneration of damaged cartilage. Although producing tissue‑engineered cartilage from marrow stem cells appeared to be a feasible method, constructing certain sub‑types of cartilage, including elastic cartilage, remains difficult. Therefore, the present study explored the feasibility of constructing elastic cartilage by culturing bone marrow‑derived stem cells (BMSCs) in the supernatant of elastic cartilage cells to generate elastic cartilage. The elastic cartilage cells were obtained from the auricle cartilage of a newborn pig, and BMSCs were isolated from pig bone marrow aspirate. The supernatant of the chondrocytes was collected and then used to the culture BMSCs. At various time‑points, the differentiation of BMSCs was evaluated by gross view, histological examination and quantitative polymerase chain reaction. BMSCs changed from spindle‑shaped cells into polygonal cells with increasing culture time. The expression of collagen II and elastin was observed in the cells cultured in the supernatant of elastic chondrocytes, while no expression was observed in the control cells. Furthermore, the expression of collagen I and collagen X was downregulated in the cells cultured in the supernatant of elastic cartilage cells. The supernatant of elastic cartilage cells promoted the differentiation of BMSCs into elastic cartilage cells, which may be a promising method for constructing certain sub‑types of tissue‑engineered cartilage.
Isolation, identification, and comparison of cartilage stem progenitor/cells from auricular cartilage and perichondrium.
Xue Ke,Zhang Xiaodie,Qi Lin,Zhou Jia,Liu Kai
American journal of translational research
Auricular cartilage loss or defect remains a challenge to plastic surgeons, and cartilage regenerative medicine provides a novel method to solve the problem. However, ideal seeding cells seem to be the key point in the development of cartilage regeneration. Although bone marrow-mesenchymal stem cells were considered as the ideal seeding cells in cartilage regeneration, regenerative cartilage differentiated from bone marrow-mesenchymal stem cells still faces some problems. It is reported that many tissues and organs contain a certain number of adult progenitor or stem cells that can replace cells that die or restore tissues and organs after injury. Therefore, we tried to use a fibronectin differential adhesion assay to isolate cartilage stem/progenitor cells from auricular cartilage and perichondrium. Flow cytometric analysis demonstrated the two cell populations expressed mesenchyme stem cell positive surface marker. Meanwhile, the cells differentiate into osteogenic line, chondrogenic line and adipogenic line under different induction conditions. The proliferation of cartilage stem/progenitor cells derived from perichondrium was higher than cartilage stem/progenitor cells derived from auricular cartilage. In addition, there is a difference on osteogenic differentiation, chondrogenic differentiation and adipogenic differentiation between these two cell populations. In conclusion, auricular cartilage and perichondrium both contain cartilage stem/progenitor cells, which may provide an ideal seeding cells for cartilage regeneration.
Domino Flaps for Repairing of Secondary Bilateral Cleft Lip With Severe Columella Deformity.
Wei Jiao,Chiang Cheng-An,Zhou Da,Li Qingfeng,Liu Kai
The Journal of craniofacial surgery
The patients with secondary bilateral cleft lip and nasal deformity always suffer from a serious part of the horizontal and vertical soft tissue deficiencies in the upper lip and nose, especially the columella. Normally, the Abbe flap is used to increase the soft tissue volume of upper lip. However, how to reconstruct the sever columella deficient, how to make full use of the Abbe flap and the remaining soft tissue of the upper lip, are always the problems that need to be solved by plastic surgeons. Twenty-nine patients with secondary bilateral cleft lip and severe columella deficiency were simultaneously reconstructed with several local flaps like dominos that was called "Domino sliding flaps." Six of the patients had their nasal tip and dorsum or collumella augmented and modified with autologous costal cartilage at the same time. The other 23 patients underwent the autologous auricular cartilage augmentation. The patients were followed from 15 months to 50 months. No secondary deformation occurred in any of the patients. 93.1% patients were satisfied with the aesthetics postoperatively, 89.7% patients were satisfied with the incision and donor site scar. Among them, 3 of the patients got hypertrophic scar on the upper lip and were modified 1 year after their first operations. The operational technique of the domino sliding flaps could better play the role of the Abbe flap. It also gave plastic surgeons an effective method to modify the severe deformities of bilateral cleft lip in 1 stage.
Exosomes derived from mature chondrocytes facilitate subcutaneous stable ectopic chondrogenesis of cartilage progenitor cells.
Chen Yahong,Xue Ke,Zhang Xiaodie,Zheng Zhiwei,Liu Kai
Stem cell research & therapy
BACKGROUND:Developing cartilage constructed with the appropriate matrix composition and persistent chondrogenesis remains an enduring challenge in cartilage defects. Cartilage progenitor cell (CPC)-based tissue engineering has attracted recent attention because of its strong chondrogenic differentiation capacity. However, due to the lack of a suitable chondrogenic niche, the clinical application of CPC-regenerated cartilage in the subcutaneous environment remains a challenge. In this study, exosomes derived from chondrocytes (CC-Exos) were used to provide the CPC constructs with a cartilage signal in subcutaneous environments for efficient ectopic cartilage regeneration. METHODS:Rabbit CPC-alginate constructs were prepared and implanted subcutaneously in nude mice. CC-Exos were injected into the constructs at the same dose (30 μg exosomes per 100 μL injection) after surgery and thereafter weekly for a period of 12 weeks. Exosomes derived from bone mesenchymal stem cells (BMSC-Exos) were used as the positive control. The mice in the negative control were administered with the same volume of PBS. At 4 and 12 weeks after implantation, the potential of CC-Exos and BMSC-Exos to promote chondrogenesis and stability of cartilage tissue in a subcutaneous environment were analyzed by histology, immunostaining, and protein analysis. The influences of BMSC-Exos and CC-Exos on chondrogenesis and angiogenic characteristics in vitro were assessed via coculturing with CPCs and human umbilical vein endothelial cells. RESULTS:The CC-Exos injection increased collagen deposition and minimized vascular ingrowth in engineered constructs, which efficiently and reproducibly developed into cartilage. The generated cartilage was phenotypically stable with minimal hypertrophy and vessel ingrowth up to 12 weeks, while the cartilage formed with BMSC-Exos was characterized by hypertrophic differentiation accompanied by vascular ingrowth. In vitro experiments indicated that CC-Exos stimulated CPCs proliferation and increased expression of chondrogenesis markers while inhibiting angiogenesis. CONCLUSIONS:These findings suggest that the novel CC-Exos provides the preferable niche in directing stable ectopic chondrogenesis of CPCs. The use of CC-Exos may represent an off-the-shelf and cell-free therapeutic approach for promoting cartilage regeneration in the subcutaneous environment.
Midline Volume Filler Injection for Facial Rejuvenation and Contouring in Asians.
Xiong Zhezhen,Jiang Zhengzheng,Liu Kai
Aesthetic plastic surgery
BACKGROUND:Hyaluronic acid (HA) injection is popular in cosmetic surgery because of its minimal invasion and immediate recovery. However, few injection techniques are specifically designed for Asians, considering their aesthetic and structural differences from those of Caucasians. METHODS:The midline volume injection technique was performed on 37 Asian females and 3 males at sites including the forehead, glabella, nasal dorsum, nasal base and chin. The treatment focused on improving the facial profile projection. 3D image data were collected, and facial aesthetic angles were calculated. Volume changes in facial parts were analyzed before injection and during follow-up. Patient satisfaction was assessed using the visual analog scale (VAS). RESULTS:The average injected filler volume was 6.14 ± 2.82 ml. The treatment optimized the facial contour and produced instant facial rejuvenation. On 3D images, the average midline volume increased by 6.02 ± 2.23 ml. An average volume decrease of 3.92 ± 1.94 ml was observed in the cheeks and was positively related to the increased midline volume. The average nasolabial and nasal facial angles were increased from 100.00 ± 7.37° to 107.93 ± 9.01° and 28.78 ± 3.28° to 31.78 ± 2.97°, while the mentolabial angle was decreased from 146.55 ± 7.83° to 141.13 ± 6.23°. The mean VAS scores were 2.28 ± 1.27 immediately after injection and 2.56 ± 1.20 after 6 months. CONCLUSIONS:The midline volume injection technique of HA fillers is suitable for Asians. It greatly enhances the facial profile, as illustrated by volume changes in 3D images, leading to a more attractive facial appearance. LEVEL OF EVIDENCE V:This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.
Silicate bioceramics enhanced vascularization and osteogenesis through stimulating interactions between endothelia cells and bone marrow stromal cells.
Li Haiyan,Xue Ke,Kong Ni,Liu Kai,Chang Jiang
The facts that biomaterials affect the behavior of single type of cells have been widely accepted. However, the effects of biomaterials on cell-cell interactions have rarely been reported. Bone tissue engineering involves osteoblastic cells (OCs), endothelial cells (ECs) and the interactions between OCs and ECs. It has been reported that silicate biomaterials can stimulate osteogenic differentiation of OCs and vascularization of ECs. However, the effects of silicate biomaterials on the interactions between ECs and OCs during vascularization and osteogenesis have not been reported, which are critical for bone tissue regeneration in vivo. Therefore, this study aimed to investigate the effects of calcium silicate (CS) bioceramics on interactions between human umbilical vein endothelial cells (HUVECs) and human bone marrow stromal cells (HBMSCs) and on stimulation of vascularization and osteogenesis in vivo through combining co-cultures with CS containing scaffolds. Specifically, the effects of CS on the angiogenic growth factor VEGF, osteogenic growth factor BMP-2 and the cross-talks between VEGF and BMP-2 in the co-culture system were elucidated. Results showed that CS stimulated co-cultured HBMSCs (co-HBMSCs) to express VEGF and the VEGF activated its receptor KDR on co-cultured HUVECs (co-HUVECs), which was also up-regulated by CS. Then, BMP-2 and nitric oxide expression from the co-HUVECs were stimulated by CS and the former stimulated osteogenic differentiation of co-HBMSCs while the latter stimulated vascularization of co-HVUECs. Finally, the poly(lactic-co-glycolic acid)/CS composite scaffolds with the co-cultured HBMSCs and HUVECs significantly enhanced vascularization and osteogenic differentiation in vitro and in vivo, which indicates that it is a promising way to enhance bone regeneration by combining scaffolds containing silicate bioceramics and co-cultures of ECs and OCs.
Small Diameter Blood Vessels Bioengineered From Human Adipose-derived Stem Cells.
Zhou Renpeng,Zhu Lei,Fu Shibo,Qian Yunliang,Wang Danru,Wang Chen
Bioengineering of small-diameter blood vessels offers a promising approach to reduce the morbidity associated with coronary artery and peripheral vascular disease. The aim of this study was to construct a two-layered small-diameter blood vessel using smooth muscle cells (SMCs) and endothelial cells (ECs) differentiated from human adipose-derived stem cells (hASCs). The outer layer was constructed with biodegradable polycaprolactone (PCL)-gelatin mesh seeded with SMCs, and this complex was then rolled around a silicone tube under pulsatile stimulation. After incubation for 6 to 8 weeks, the PCL-gelatin degraded and the luminal supporting silicone tube was removed. The smooth muscle layer was subsequently lined with ECs differentiated from hASCs after stimulation with VEGF and BMP4 in combination hypoxia. The phenotype of differentiated SMCs and ECs, and the cytotoxicity of the scaffold and biomechanical assessment were analyzed. Our results demonstrated that the two-layered bioengineered vessels exhibited biomechanical properties similar to normal human saphenous veins (HSV). Therefore, hASCs provide SMCs and ECs for bioengineering of small-diameter blood vessels.