ObjectiveTo summarize the research progress of several three-dimensional (3-D) printing scaffold materials in bone tissue engineering. MethodThe recent domestic and international articles about 3-D printing scaffold materials were reviewed and summarized. ResultsCompared with conventional manufacturing methods, 3-D printing has distinctive advantages, such as enhancing the controllability of the structure and increasing the productivity. In addition to the traditional metal and ceramic scaffolds, 3-D printing scaffolds carrying seeding cells and tissue factors as well as scaffolds filling particular drugs for special need have been paid more and more attention. ConclusionsThe development of 3-D printing porous scaffolds have revealed new perspectives in bone repairing. But it is still at the initial stage, more basic and clinical researches are still needed.
Objective To observe the biocompatibility of the acellular corneal stroma materials prepared by three different methods. Methods Three different serial digestion methods were used to produce the acellular corneal stroma materials. The biocompatibility of the materials was investigated by the cell seeding and the materials were implanted into the rabbit corneal stroma layer. Results The cells in the materials 1 and 2 were not decellularized completely. The rabbit corneal fibroblasts died on the materials 1 and 2 after the cell seeding for 3-4 days. An obvious rejection could be observed after the implantation. The cells in material 3 were decellularized completely and the collagen fibers or elastic fibers were reserved integrally，showing a typical three-dimensional net work. The rabbit corneal fibroblasts could expand on the materials in vitro. No obvious rejection could be observed and the materials were gradually absorbed. Conclusion The acellular porcine cornea stroma materials prepared by trypsin-Dnase-Rnase are suitable for reconstruction of the tissue engineered cornea.
The engineered heart tissues (EHTs) present a promising alternative to current materials for native myocardial tissue due to the unique characteristics. However, until now, the clinical application of EHTs is limited by a serial of practical problems yet. Generally, the challenges need to further optimize include biomaterials, cell sources, and strategies of revascularization or establishment of EHTs. This review focuses on the newly progress on these aspects to encourage the emergence of novel EHTs that can meet clinic requirement properly.
【Abstract】 Objective To compare the effect of PLGA and collagen sponge combined with rhBMP-2 on repairing ofarticular cartilage defect in rabbits respectively. Methods PLGA and collagen sponge were made into cyl inders which were 4 mm in diameter and 3 mm in thickness, and compounded with rhBMP-2 (0.5 mg). Defect 4 mm in diameter were made in both of femoral condyles of 24 two-month-old New Zealand white rabbits. The defects in right 18 knees were treated with PLGA/rhBMP-2 composites (experimental group 1), and the left 18 knees were treated with collagen sponge/rhBMP-2 composites (experimental group 2), the other 12 knees were left untreated as control group. At 4, 12 and 24 weeks after operation, the animals were sacrificed and the newly formed tissues were observed macroscopically and microscopically, graded histologically and analyzed statistically. Results From the results of macroscopical and microscopical observation, in the experimental group 1, the defects were filled with smooth and translucent cartilage; while in the experimental group 2, the white translucent tissues did notfill the defects completely; and in the two experimental groups, the new cartilage tissues demarcated from the surrounding cartilage,chondrocytes distributed uniformly but without direction; a l ittle fibrous tissue formed in the control group 4 weeks postoperatively. In the experimental group 1, the defects were filled completely with white, smooth and translucent cartilage tissue without clear l imit with normal cartilage; while in the experimental group 2, white translucent tissues formed, the boundary still could be recognized; in the two experimental groups, the thickness was similar to that of the normal cartilage; the cells paralleled to articular surface in the surface layer, but in the deep layer, the cells distributed confusedly, the staining of matrix was positive but a l ittle weak; subchondral bone and tide mark recovered and the new tissue finely incorporated with normal cartilage;however, in the control group, there was a l ittle of discontinuous fibrous tissue, chondrocytes maldistributed in the border andthe bottom of the defects 12 weeks postoperatively. In the experimental group 1, white translucent cartilage tissues formed, the boundary disappeared; in the experimental group 2, the color and the qual ity of new cartilage were similar to those of 12 weeks; in the two experimental groups, the thickness of the new cartilage, which appeared smooth, was similar to that of the normal cartilage, the chondrocytes arranged uniformly but confusedly; the staining of matrix was positive and subchondral bone and tide mark recovered, the new tissue finely incorporated with normal cartilage; in the control group, a layer of discontinuous fibrous tissue formed in the bottom of the defects 24 weeks postoperatively. Results of histological grade showed that there were significantdifference between experimental group (1 and 2) and control group at any time point (P lt; 0.01); the scores of 12 weeks and 24 weeks in experimental group 1 and 2 had a significant difference compared with that of 4 weeks (P lt; 0.01), there was no significant difference between 12 weeks and 24 weeks (P gt; 0.05), and there were no significant difference between the two experimental groups at the same time point (P gt; 0.05). Conclusion Both PLGA and collagen sponge as a carrier compounded with rhBMP-2 can repair articular cartilage defects.
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.
Objective To investigate the feasibil ity of using thermo-sensitive chitosan hydrogen as a scaffold to construct tissue engineered injectable nucleus pulposus (NP). Methods Three-month-old neonatal New Zealand rabbits (male or female) weighing 150-200 g were selected to isolate and culture NP cells. The thermo-sensitive chitosan hydrogel scaffold wasmade of chitosan, disodium β-glycerophosphate and hydroxyethyl cellulose. Its physical properties and gross condition were observed. The tissue engineered NP was constructed by compounding the scaffold and rabbit NP cells. Then, the viabil ity of NP cells in the chitosan hydrogel was observed 2 days after compound culture and the growth condition of NP cells on the scaffold was observed by SEM 7 days after compound culture. NP cells went through histology and immunohistochemistry detection and their secretion of aggrecan and expression of Col II mRNA were analyzed by RT-PCR 21 days after compound culture. Results The thermo-sensitive chitosan hydrogel was l iquid at room temperature and sol idified into gel at 37 (15 minutes) due to crossl inking reaction. Acridine orange-propidiumiodide staining showed that the viabil ity rate of NP cells in chitosan hydrogel was above 90%. Scanning electron microscope observation demonstrated that the NP cells were distributed in the reticulate scaffold, with ECM on their surfaces. The results of HE, toluidine blue, safranin O and histology and immunohistochemistry staining confirmed that the NP cells in chitosan hydrogel were capable of producing ECM. RT-PCR results showed that the secretion of Col II and aggrecan mRNA in NP cells cultured three-dimensionally by chitosan hydrogen scaffold were 0.631 ± 0.064 and 0.832 ± 0.052, respectively,showing more strengths of producing matrix than that of monolayer culture (0.528 ± 0.039, 0.773 ± 0.046) with a significant difference (P lt; 0.05). Conclusion With good cellular compatibilities, the thermo-sensitive chitosan hydrogel makes it possible for NP cells to maintain their normal morphology and secretion after compound culture, and may be a potential NP cells carrier for tissue engineered NP.
Objective To study the cellular biocompatibility, adhesion and proliferation of endothelial outgrowth cells (EOCs) isolated and expanded from rabbit peripheral blood cultured with aligned poly-L-lactic acid (PLLA) nanofibrous scaffolds in vitro so as to provide a basis for the applications of scaffolds biomaterials in tissue repair. Methods Nanofibrous scaffolds of PLLA by electrostatic spinning were modified by hypothermal plasmas body and type Ⅰ collagen was coated onto the materials physically. In vitro, EOCs were cultured on the modified PLLA scaffold. Adhesion and proliferation were surveyed and morphological changes and biocompatibility of seeding cells on PLLA scaffold were observed by growth curves of the cells, fluorescent microscope and scanning electron microscope respectively. Results Fibers with diameters ranging from 300 nm to 400 nm were included in the nanofibrous scaffolds, whose porosities were more than 90%. Absorbance (A) of each scaffold increased gradually after EOCs grew in the absence or presence of random, aligned, or super-aligned PLLA nanofibrous scaffold. Although there was no detectable effect of the random PLLA scaffold on the growth EOCs (Pgt;0.05), both aligned and super-aligned PLLA nanofibrous scaffold had significantly enhanced their growth since the 5th day (P＜0.05). The rates of adhesion in both aligned and super-aligned PLLA nanofibrous scaffold were significantly higher than those of random PLLA scaffold after 12 h and 24 h incubation (P＜0.01). The rates of proliferation after 1 d, 3 d and 7 d incubation in aligned and super-aligned PLLA nanofibrous scaffold were significantly higher than those of random PLLA nanofibrous scaffold (P＜0.05, P＜0.01). EOCs grew well with PLLA scaffold, yet confused and disorderly in random nanofibers. EOCs could attach, extend and proliferate following fibrous orientation in aligned and super-aligned PLLA nanofibrous scaffold, in majority of the fibers were oriented along the longitudinal axis so that a unique aligned topography was formed. Especially super-aligned PLLA nanofibrous had advantageous to keep well on cell morphology. Conclusion EOCs are ideal seeding cells for tissue engineering. EOCs can be adhered well to aligned and super-aligned PLLA nanofibrous scaffold and proliferate, keep well on cell morphology. So this type of PLLA nanofibrous scallfold is proposed to be an optimal candidate material for EOCs transplantation in tissue repair.
ObjectiveTo study the preparation method of acellular dermal matrix (ADM) for cartilage tissue engineering and analyze its biocompatibility. MethodsThe dermal tissues of the calf back were harvested, and decelluarized with 0.5% SDS, and the ADM was reconstructed with 0.5% trypsin, cross-linked with formaldehyde, and modified with 0.5% chondroitin sulfate which can promote the proliferation of chondrocytes. And the porosity, cytotoxicity, and biocompatibility were determined. Co-cultured 2nd passage chondrocytes and bone marrow stromal cells in a proportion of 3 to 7 were used as seed cells. The cells were seeded on ADM (experimental group) for 48 hours to observe the cell adhesion. The expressions of mRNA and protein of collagen type Ⅱ were tested by RT-PCR and Western blot methods, respectively. And the expressions were compared between the cells seeded on the scaffold and cultured in monolayer (control group). ResultsAfter modification of 0.5% trypsin, the surface of ADM was smooth and had uniform pores; the porosity (85.4%±2.8%) was significantly higher than that without modification (72.8%±5.8%) (t=-4.384, P=0.005). The cell toxicity was grade 1, which accords to the requirements for cartilage tissue engineering scaffolds. With time passing, the number of inflammatory cells decreased after implanted in the back of the rats (P<0.05). The scanning electron microscope observation showed that lots of seed cells adhered to the scaffold, the cells were well stacked, displaying surface microvilli and secretion. The expressions of mRNA and protein of collagen type Ⅱ were not significantly different between experimental and control groups (t=1.265, P=0.235;t=0.935, P=0.372). ConclusionThe ADM prepared by acellular treatment, reconstruction, cross-linking, and modification shows perfect characters. And the seed cells maintain chondrogenic phenotype on the scaffold. So it is a proper choice for cartilage tissue engineering.
Objective To establish a scaffold model from heterogeneoussmall blood vessels. Methods Caudal arteries from 34 Wistar rats( average length 12.08±1.69 cm) were made into acellular blood vessel scaffolds. Some scaffoldswere observed by electron microscope, and others were transplanted to the cut ends of ear central arteries of male Japanese big ear white rabbits. Results Average external diameter was 0.74±0.08 mm in proximal, and 0.55±0.08 mm in distal end of rat caudal arteries. The small blood vessel scaffolds had shin wall whichwas white and soft, composed of fibrous tissues without cells. On the intima surface the fibrous tissues were arrayed densely in a grid-like pattern. After transplantation, the blood flow was reserved, and kept flowing freely in 24 hours. The pulsation of the transplanted artery was accessible and no blood leakage wasfound.Conclusion The natural scaffolds are composed of fibrous tissues, and can sustain the artery pulse pressure for 24 hours. It is better to suture the blood vessels by sleeve anastomosis.
Objective To study the mechanism of ectopic osteogenesis of nacre/Polylactic acid (N/P) artificial bone combined with allogenic osteoblasts, and to explore the possibility as a scaffold material of bone tissue engineering. Methods The allogenic- osteoblasts seeded onto N/P artificial bone were co-cultured in vivo 1 week.The N/P artificial bone with allogenic osteoblasts were implanted subcutaneously into the left back sites of the New Zealand white rabbits in the experimental group and the simple N/P artificial bone into the right ones in the control group. The complexes were harvested and examined by gross observation, histologic analysis and immunohistochemical investigation 2, 4 and 8 weeks after implantation respectively.Results In experimental group, the osteoid formed after 4 weeks, and the mature bone tissue withbone medullary cavities formed after 8 weeks; but in control group there was nonew bone formation instead of abundant fibrous tissue after 4 weeks, and more fibrous tissue after 8 weeks.Conclusion N/P artificial bone can be used as an optical scaffold material of bone tissue engineering.