Objective To develop a new method for a tissue engineered vascular graft by combining endothelial cells and an acelluarized allogenic matrix. Methods Acellularized matrix tubes were obtained by a 0.1% trypsin and 0 02% EDTA solution for 24 hours and 1% Triton X 100 for 176 hours, respectively. Endothelial cells were isolated from alloaorta and expanded in vitro. Finally, the inner surface of acellularized matrix was reseeded with endothelial cells. Acellularity and reseeding were analysed by light microscopy and scanning electron microscopy. Results The acellularization procedure resulted in an almost complete removal of the original cells and the loose three-dimensional (3D) matrix. The acellular matrix could be reseeded with expanded endothelial cells in vitro, and endothelial cells had the potential of spread and proliferation. Conclusion Acellular matrix produces by Tritoon X-100 and trypsin possesses satisfactory biocompatibility for allogenic endothelial cell. Vascular grafts can be generated in vitro by a combination of endothelial cells and allogenic acelluarized matrix.
OBJECTIVE To investigate the methods to fabricate repair materials of tissue engineered peripheral nerve with bioactivity of Schwann cells (SC). METHODS 1. The materials were made by dry-wet spinning process to fabricate PLA hollow fiber canal with external diameter of 2.3 mm, internal diameter of 1.9 mm, thickness of 0.4 mm, pore size of 20 to 40 microns, pore ratio of 70% and non-spinning fiber net with pore size of 100 to 200 microns, pore ratio of 85%. 2. SC were implanted into excellular matrix (ECM) gel to observe the growth of SC. 3. SC/ECM complex were implanted into non-spinning PLA fiber net to observe the growth of SC. 4. SC, SC/ECM and SC/ECM/PLA were implanted into PLA hollow fiber canal to bridge 10 mm defect of rat sciatic nerve. RESULTS 1. SC were recovered bipolar shape at 1 day after implantation, and could be survived 14 days in ECM gel. 2. After SC/ECM complex was implanted into PLA net, most of SC were retained in the pore of PLA net with the formation of ECM gel. SC could be adhered and grown on PLA fiber. 3. Most of SC in ECM gel could be survived to 21 days after transplantation. Survival cell numbers of SC/ECM and SC/ECM/PLA groups were obviously higher than SC suspension group. CONCLUSION Non-spinning PLA porous biodegradable materials with ECM is benefit for SC to be adhered and grown.
ObjectiveTo study the feasibility of acellular matrix materials prepared from deer antler cartilage and its biological compatibility so as to search for a new member of the extracellular matrix family for cartilage regeneration. MethodsThe deer antler mesenchymal (M) layer tissue was harvested and treated through decellular process to prepare M layer acellular matrix; histologic observation and detection of M layer acellular matrix DNA content were carried out. The antler stem cells [antlerogenic periosteum (AP) cells] at 2nd passage were labelled by fluorescent stains and by PKH26. Subsequently, the M layer acellular matrix and the AP cells at 2nd passage were co-cultured for 7 days; then the samples were transplanted into nude mice to study the tissue compatibility of M layer acellular matrix in the living animals. ResultsHE and DAPI staining confirmed that the M layer acellular matrix did not contain nucleus; the DNA content of the M layer acellular matrix was (19.367±5.254) ng/mg, which was significantly lower than that of the normal M layer tissue [(3 805.500±519.119) ng/mg](t=12.630, P=0.000). In vitro co-culture experiments showed that AP cells could adhere to or even embedded in the M layer acellular matrix. Nude mice transplantation experiments showed that the introduced AP cells could proliferate and induce angiogenesis in the M layer acellular matrix. ConclusionThe deer antler cartilage acellular matrix is successfully prepared. The M layer acellular matrix is suitable for adhesion and proliferation of AP cells in vitro and in vivo, and it has the function of stimulating angiogenesis. This model for deer antler cartilage acellular matrix can be applied in cartilage tissue engineering in the future.
Objective To observe the expressions of extracellular matrix metalloproteinase inducer (EMMPRIN) and matrix metalloproteinase 9 (MMP-9) around the prosthesis, and to study the relationship between the expressions of EMMPRIN and MMP-9 and osteolysis around prosthesis. Methods Interface tissues were obtained at three Delee-Charnley acetabular sections and seven Gruen femur sections from 8 cases (8 hips) undergoing revision after total hip arthroplasty between February 2010 and January 2012, and were divided into osteolysis group and non-osteolysis group based on preoperative X-ray film and intraoperative observation; the tissues from another 8 patients with osteoarthritis undergoing total hip arthroplasty as the control group. The immunohistochemical staining and RT-PCR assays were used to determine the expressions of EMMPRIN and MMP- 9. The correlation between the positive cells and the severity of osteolysis were analyzed and compared. Results Histological examination showed that many macrophages, multinucleated giant cells assembled in the membrane of osteolysis zone, but many fibroblasts and synovial cells in non-osteolysis zones. EMMPRIN and MMP- 9 positive cells and gene expressions were observed in every group. The percentage of positive cells and gene expression of EMMPRIN and MMP-9 in osteolysis group were significantly higher than those in non-osteolysis and control groups (P lt; 0.05), but no significant difference was found between non-osteolysis group and control group (P gt; 0.05). The percentage of positive cells of EMMPRIN in zone III of acetabular was higher than that in zone I and zone II of revision hip (P lt; 0.05), but no significant difference between zone I and zone II (P gt; 0.05). The percentage of positive cells of MMP-9 in zone I and zone III was significantly higher than that in zone II of revision hip (P lt; 0.05), but no significant difference between zone I and zone III (P gt; 0.05). The expression of EMMPRIN from high to low in order was zones 1, 7, 4, 2, 3, 5, and 6 at femur; the values of zones 1, 7, and 4 were significantly higher than those of zones 2, 3, 5, and 6 (P lt; 0.05), but no significant difference among zones 1, 7, and 4, and among zones 2, 3, 5, and 6 (P gt; 0.05). The expression of MMP-9 from high to low in order was zones 1, 7, 4, 2, 3, 6, and 5 at femur; the values of zones 1 and 7 were significantly higher than those of zones 4, 2, 3, 6, and 5 (P lt; 0.05), and the values of zones 4 and 2 were significantly higher than those of zones 3, 6, and 5 (P lt; 0.05), but no significant difference between zone 1 and zone 7, between zone 4 and zone 2, and among zones 3, 5, and 6 (P gt; 0.05). Conclusion The expressions of EMMPRIN and MMP-9 have certain coherence. The over-expressions of EMMPRIN and MMP-9 may be one of the key points of inhibiting bone reconstruction and bone resorption at bone-implant interface under the stimulation of wear debris.
ObjectiveTo review the properties of bio-derived hydrogels and their application and research progress in tissue engineering. MethodsThe literature concerning the biol-derived hydrogels was extensively reviewed and analyzed. ResultsBio-derived hydrogels can be divided into single-component hydrogels (collagen,hyaluronic acid,chitosan,alginate,silk fibroin,etc.) and multi-component hydrogels[Matrigel,the extract of extracellular matrix (ECM),and decellularized ECM].They have favorable biocompatibility and bioactivity because they are mostly extracted from the ECM of biological tissue.Among them,hydrogels derived from decellularized ECM,whose composition and structure are more in line with the requirements of bionics,have incomparable advantages and prospects.This kind of scaffold is the closest to the natural environment of the cell growth. ConclusionBio-derived hydrogels have been widely used in tissue engineering research.Although there still exist many problems,such as the poor mechanical properties,rapid degradation,the immunogenicity or safety,vascularization,sterilization methods,and so on,with the deep-going study of optimization mechanism,desirable bio-derived hydrogels could be obtained,and thus be applied to clinical application.
ObjectiveTo summarize recent progress in adipose tissue acting as a more efficient and ideal therapy to facilitate wound repair and evaluate the therapeutic values of adipose tissue.MethodsThe related literature about adipose tissue for wound healing in recent years was reviewed and analyzed.ResultsEnormous studies focus on the capacity of adipose tissue to accelerate wound healing including cellular components, extracellular matrix, and paracrine signaling have been investigated.ConclusionAdipose tissue has generated great interest in recent years because of unique advantages such as abundant and accessible source, thriven potential to enhance the regeneration and repair of damaged tissue. However, there is still a need to explore the mechanism that adipose tissue regulates cellular function and tissue regeneration in order to facilitate clinical application of adipose tissue in wound healing.
【Abstract】Objective To investigate the expression of extracellular matrix metalloproteinase inducer(EMMPRIN),matrix metalloproteinase-1(MMP1),MMP9,tissue inhibitors of metalloproteinase-1(TIMP1) and the mast cell count (MCC) and to detect their clinicopathologic significance and relationship in pancreatic cancer tissues. Methods Immunohistochemical method of avidin-biotin complex was used for those 5 targets on the routinely paraffinembedded sections of surgical resected specimen of 51 cases with pancreatic carcinoma. Results The positive rates of EMMPRIN,MMP1,MMP9 and TIMP1 were 56.9%,54.9%,60.8% and 49.0% and its scoring were 2.5±1.5,2.3±1.9,2.4±1.6 and 1.9±1.6 respectively. The mean of MCC was (16.1±6.8)/HP in total cases. The positive rates or scorings of EMMPRIN,MMP1,MMP9 and MCC were significantly lower in high differentiated or without-metastatic cases than in low differentiated or with-metastatic ones(P<0.05 or P<0.01), and those targets (except MCC and scoring of MMP9) of middle differentiated ones were lower than those of low differentiated while that of TIMP1 was opposite(P<0.01). The MCC showed significantly higher in the positive cases of EMMPRIN, MMP1 and MMP9 or negative cases of TIMP1 than in the negative ones of EMMPRIN, MMP1 and MMP9 or positive ones of TIMP1. The closely positive correlations were found among the MCC and the scoring of EMMPRIN, MMP1 and MMP9. The closely negative correlations existed among the scoring of TIMP1 and the other four targets.Conclusion The MCC and the expressions of EMMPRIN, MMP1, MMP9 and TIMP1 might be important biological markers for reflecting the progression and the prognosis of pancreatic carcinoma. They might have co-regulated effects on the potentials of invasion and metastasis of pancreatic carcinoma or other malignant lesions.
Objective To investigate pathogenesis and therapeutic prospect of abdominal aortic aneurysm (AAA). Methods Relevant literatures about pathogenesis and ways of treatment for AAA in recent years were reviewed. Results The formation of AAA are associated with heredity, anatomy, environment and biochemistry and other factors. All factors influence and interact with each other. The metabolic disequilibrium of aortic intermediate extracellular matrix plays an important role in the pathogenesis of AAA. The main reasons for the formation of AAA may be the increase of activity of matrix metalloproteinases and the disequilibrium of genetic expressions of elastin and collagen. The therapy of AAA includes surgical and medical treatment. The methods of medical treatment are still in the process of exploration and research. Conclusion The formation of AAA is a synergistical result of multiple factors, and medical treatment is an important supplement of surgical treatment.
OBJECTIVE: To review the role of matrix metalloproteinase-1 (MMP-1) in the course of healing in wounded skin. METHODS: The recent literatures on MMP-1 in skin wound repair were reviewed, which gave the insight into the local effect of MMP-1 during re-epithelialization. RESULTS: Following injury, basal keratinocytes, moving from the wound edge and interact with dermal matrix proteins in the wound bed, were induced to express MMP-1 in a specific space-time pattern. MMP-1 cleaved the collagen, thereby altering its structure and affinity by which the keratinocytes binded it. MMP-1 served a beneficial role in wound healing by facilitating the proliferation and movement of keratinocytes over the collagen-rich wound bed during re-epithelialization. CONCLUSION: MMP-1 expression of migrating keratinocytes directly influences the re-epithelialization during the course of healing of the wounded skin.
Objective To review the research progress of promoting the bone formation at early stage by components of the extracellular matrix (ECM). Methods Recent literature concerning the influence of these components on new bone formation and bone/implant contact was extensively reviewed and summarized. Results Coating of titanium or hydroxyapatite implants with organic components of the ECM (such as collagen type I, chondroitin sulfate, and Arg-Gly-Asp peptide) offers great potential to improve new bone formation and enhance bone/implant contact, which in turn will shorten recovery time and improve implant stability. Conclusion The increasing knowledge about the role of the ECM for recruitment, proliferation, differentiation of cells, and regeneration of tissue will eventually deal to the creating of an artificial ECM on the implant that could allow a defined adjustment of the required properties to support the healing process.