Collagen Composite Scaffold Research Articles

3D‐Printed and Recombinant Spider Silk Particle Reinforced Collagen Composite Scaffolds for Soft Tissue Engineering

AbstractCollagen is one main component of the extracellular matrix (ECM) in natural tissues and is, therefore, well suited as a biomaterial for tissue engineering. In this study, a method is presented to 3D‐bioprint collagen into a precipitation bath comprising recombinantly produced spider silk protein eADF4(C16) yielding a composite with excellent mechanical properties. The spider silk precipitation bath induced assembly of the collagen into fibrils, and subsequent addition of potassium phosphate buffer lead to the formation of silk particles and stabilization of the collagen fibrils. The produced collagen‐silk composite scaffolds show an internal structure of homogeneously distributed and interacting collagen fibrils and spider silk particles with significantly better mechanical properties compared to plain collagen scaffolds. Further, enzymatic degradation assays of the scaffolds over a 7‐day period show higher stability of the collagen‐silk scaffolds compared to plain collagen scaffolds in the presence of wound proteases. Using the spider silk variant eADF4(C16‐RGD) further increases compressive stress and elastic modulus compared to that of the unmodified variant. Finally, it is shown that the unique collagen‐spider silk composite scaffolds comprising the cell‐binding domains of collagen and the RGD sequence in the spider silk variant represent a promising material for soft tissue regeneration.

Reconstructing Critical-Sized Mandibular Defects in a Rabbit Model: Enhancing Angiogenesis and Facilitating Bone Regeneration via a Cell-Loaded 3D-Printed Hydrogel-Ceramic Scaffold Application.

In this study, we propose a spatially patterned 3D-printed nanohydroxyapatite (nHA)/beta-tricalcium phosphate (β-TCP)/collagen composite scaffold incorporating human dental pulp-derived mesenchymal stem cells (hDP-MSCs) for bone regeneration in critical-sized defects. We investigated angiogenesis and osteogenesis in a rabbit critical-sized mandibular defect model treated with this engineered construct. The critical and synergistic role of collagen coating and incorporation of stem cells in the regeneration process was confirmed by including a cell-free uncoated 3D-printed nHA/β-TCP scaffold, a stem cell-loaded 3D-printed nHA/β-TCP scaffold, and a cell-free collagen-coated 3D-printed nHA/β-TCP scaffold in the experimental design, in addition to an empty defect. Posteuthanasia evaluations through X-ray analysis, histological assessments, immunohistochemistry staining, histomorphometry, and reverse transcription-polymerase chain reaction (RT-PCR) suggest the formation of substantial woven and lamellar bone in the cell-loaded collagen-coated 3D-printed nHA/β-TCP scaffolds. Histomorphometric analysis demonstrated a significant increase in osteoblasts, osteocytes, osteoclasts, bone area, and vascularization compared to that observed in the control group. Conversely, a significant decrease in fibroblasts/fibrocytes and connective tissue was observed in this group compared to that in the control group. RT-PCR indicated a significant upregulation in the expression of osteogenesis-related genes, including BMP2, ALPL, SOX9, Runx2, and SPP1. The findings suggest that the hDP-MSC-loaded 3D-printed nHA/β-TCP/collagen composite scaffold is promising for bone regeneration in critical-sized defects.

Biomimetic collagen composite matrix-hydroxyapatite scaffold induce bone regeneration in critical size cranial defects

Although biomimetic scaffolds have been constructed to repair critical-size bone defects, the creation of composite scaffolds that can mimic the anisotropy of natural bone remains a major challenge. Herein, we have for the first time developed a biomimetic collagen composite matrix-hydroxyapatite (CCMH) scaffold, which highly resembles both the composition and the anisotropy hierarchical structure of natural bone. The CCMH scaffold was created through unidirectional freeze-casting and in situ peristaltic mineralization, resulting in uniformly distributed nano-hydroxyapatite on the porous collagen composite matrix (CCM) scaffold. The CCMH scaffold finely mimics the composition and anisotropic channel-like morphology of native bone, which exhibits high biocompatibility, excellent cellular activity and extraordinary osteogenic differentiation capability. Magnetic resonance imaging (MRI), micro-computed tomography (micro-CT) and histological analysis of rat models demonstrated that the CCMH scaffold significantly enhanced new bone formation and accelerated bone regeneration. The innovative unidirectional freezing-in situ peristaltic mineralization approach offers a robust technique for building biomimetic three-dimensional porous scaffolds with excellent biocompatibility and osteoinductivity, which show significant promise for use in bone repair and regeneration.

Development, Characterization, and Antibacterial Analysis of the Selenium-Doped Bio-Glass-Collagen-Gelatin Composite Scaffold for Guided Bone Regeneration.

Background Guided bone regeneration (GBR) is an often-used technique to aid the successful placement of dental implants in sites with deficient bone. The search for the ideal GBR membrane with bioactive components improving the regenerative outcomes is still on. In this study, a novel composite GBR membrane was developed using selenium-doped bio-glass, collagen, and gelatin. It was further characterized for surface, chemical, biocompatibility, and antibacterial properties. Methodology Selenium-doped bio-glass was prepared using the sol-gel method. The membrane was fabricated using an equal ratio of collagen and gelatin mixed with 1% selenium-doped bio-glass. The solution was poured to obtain a thin layer of the material which was lyophilized to obtain the final GBR membrane. The membrane was analyzed with scanning electron microscopy, energy dispersive X-ray (EDX) analysis, attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), zebrafish cytotoxicity test, and antibacterial assay. Results The membrane revealed good surface roughness with lamellar and fibrillar arrangement with a minute granular surface ideal for cell attachment and proliferation. The EDX analysis revealed the presence of carbon, oxygen, and nitrogen as predominant components with trace amounts of calcium, phosphorus, silica, and selenium. Fourier transform infrared spectroscopy analysis also proved the presence of collagen, gelatin, and bio-glass. The membrane revealed excellent biocompatibility with zebrafish growth at a normal rate with 90% viability maintained at 48, 72, and 96 hours and 95% viability at 120 hours. It also exhibited excellent antibacterial activity against Staphylococcus aureus and Escherichia coli with minimal growth of bacterial colonies. Conclusion The developed novel selenium bio-glass collagen and gelatin composite scaffold has a good surface and antibacterial properties along with excellent biocompatibility. Further cell line and in vivo studies should be conducted to explore its role in bone regeneration.

Effect of Calcium Silicate and β-Tricalcium Phosphate Reinforcement on the Mechanical–Biological Properties of Freeze-Dried Collagen Composite Scaffolds for Bone Tissue Engineering Applications

The development of a collagen-based composite scaffold to repair damaged bone is one of many important issues in tissue engineering. In this study, pure collagen, collagen/β-tricalcium phosphate (β-TCP), collagen/calcium silicate (CS), and collagen/β-TCP/CS scaffolds were fabricated using the freeze-drying method. The phase compositions, microstructures, and mechanical properties were characterized using X-ray diffraction, scanning electron microscopy, and a universal testing machine, respectively. In addition, cell viability was evaluated using an MTT assay. Finally, the correlations between the density, mechanical properties, and biodegradation behaviors of pore size distributions were discussed.

MXene-modified 3D printed scaffold for photothermal therapy and facilitation of oral mucosal wound reconstruction

The most prevalent oral and maxillofacial cancer is oral squamous cell carcinoma (OSCC). Patient survival is compromised by relapse due to post-operative tumor remnants and significant mucosal defects. Photothermal therapy (PTT) is used to ablate local malignancies, but the capacity of tumor cells to resist it is correlated with the overexpression of heat shock protein 70 (HSP70). In this work, PTT and tissue engineering scaffold were rationally integrated to construct a Ti3C2 MXene, collagen, silk fibroin and quercetin composite scaffold (M-CSQ scaffold), a 3D printed biomaterial that simultaneously kill OSCC cells and promote the regeneration of the mucosal defects. The M-CSQ scaffolds were prepared by cryogenic 3D printing and freeze-drying techniques with sufficient pores that allow an abundant cell migration and provide a proliferation space. Ti3C2 MXene has excellent photothermal conversion ability and stability. Quercetin targeted HSP70 to decrease its expression in OSCC cells, consequently weakening their resistance to high temperature and enhancing the effect of PTT. The M-CSQ scaffold effectively killed OSCC cells in vitro and inhibited tumor growth in vivo. In addition, the M-CSQ scaffold provided adhesion sites for Sprague-Dawley rat (SD rat) buccal mucosal fibroblasts and promoted the repair of buccal mucosal wounds.

Fabrication of hybrid povidone-iodine impregnated collagen-hydroxypropyl methylcellulose composite scaffolds for wound-healing application

Biomaterials derived from biological resources are advantageous in tissue engineering owing to their biocompatibility. This work presents the fabrication of a collagen composite scaffold in combination with hydroxypropyl methylcellulose (HPMC). Based on FTIR spectroscopic analysis, it is established that the native structure of collagen (Col) is unaltered with the addition of HPMC in making collagen-HPMC (ColH) composite scaffolds. Scanning electron microscopic (SEM) examination shows a smooth surface with interconnected pores in the prepared hybrid scaffolds. Considering the mechanical properties, collagen degradation, and biocompatibility, the ColH scaffold comprising 1:1.5 (w/w) of Col: HPMC was found to be optimum for applications such as wound healing and other soft tissue engineering applications. In order to determine drug loading ability of the ColH scaffold, Povidone-iodide (PI) was used as a model drug. PI loaded ColH exhibited controlled release of the drug. Further, the in vitro studies of the synthesized drug-loaded ColH scaffold showed that they effectively facilitated the proliferation of NIH3T3 fibroblasts cells without exhibiting toxicity. This study establishes the possibility of using ColH as an effective and advanced option for regenerative tissue engineering applications.

3D printed biocompatible graphene oxide, attapulgite, and collagen composite scaffolds for bone regeneration.

Tissue-engineered bone material is one of the effective methods to repair bone defects, but the application is restricted in clinical because of the lack of excellent scaffolds that can induce bone regeneration as well as the difficulty in making scaffolds with personalized structures. 3D printing is an emerging technology that can fabricate bespoke 3D scaffolds with precise structure. However, it is challenging to develop the scaffold materials with excellent printability, osteogenesis ability, and mechanical strength. In this study, graphene oxide (GO), attapulgite (ATP), type I collagen (Col I) and polyvinyl alcohol were used as raw materials to prepare composite scaffolds via 3D bioprinting. The composite materials showed excellent printability. The microcosmic architecture and properties was characterized by scanning electron microscopy, Fourier transform infrared and thermal gravimetric analyzer, respectively. To verify the biocompatibility of the scaffolds, the viability, proliferation and osteogenic differentiation of Bone Marrow Stromal Cells (BMSCs) on the scaffolds were assessed by CCK-8, Live/Dead staining and Real-time PCR in vitro. The composited scaffolds were then implanted into the skull defects on rat for bone regeneration. Hematoxylin-eosin staining, Masson staining and immunohistochemistry staining were carried out in vivo to evaluate the regeneration of bone tissue.The results showed that GO/ATP/COL scaffolds have been demonstrated to possess controlled porosity, water absorption, biodegradability and good apatite-mineralization ability. The scaffold consisting of 0.5% GO/ATP/COL have excellent biocompatibility and was able to promote the growth, proliferation and osteogenic differentiation of mouse BMSCs in vitro. Furthermore, the 0.5% GO/ATP/COL scaffolds were also able to promote bone regeneration of in rat skull defects. Our results illustrated that the 3D printed GO/ATP/COL composite scaffolds have good mechanical properties, excellent cytocompatibility for enhanced mouse BMSCs adhesion, proliferation, and osteogenic differentiation. All these advantages made it potential as a promising biomaterial for osteogenic reconstruction.

Chitin-hydroxyapatite-collagen composite scaffolds for bone regeneration

Bone defect is usually difficult to recover quickly, and bone scaffold transplantation is considered to be an effective method. Biomaterials have a wide range of application prospects in bone tissue repair, and the two key problems are the selection of materials and cells. The object of this study was to discuss the structural characteristics of bone scaffold materials and their effects on bone repair in vivo. The chitin-hydroxyapatite (HAP)-collagen composite scaffolds (CHCS) was prepared with epichlorohydrin (ECH) as crosslinking agent. The structure was characterized and the compressive strength, porosity, water absorbency and stability were investigated. The biocompatibility and osteogenic differentiation of CHCS in vitro were detected, and the effect of defect repair in vivo was evaluated. The results suggested that HAP not only enhanced the compressive strength of CHCS, but also promoted the formation of calcium nodules due to its bone conductivity. Histological staining showed that collagen promoted collagen deposition and new bone formation. X-ray images also indicated that CHCS transplantation accelerated bone repair. Therefore, CHCs has immense potential in bone regeneration.

Collagen nanomaterial in the recovery of motor ability of patients with peripheral nerve transection injury

Objective: The combination of neurotrophic factors and ordered collagen material can promote the regeneration of nerve fibres in the injured area, which has a good effect on the recovery of motor ability in patients with peripheral nerve transection. In this paper, the collagen composite scaffolds were used to repair peripheral nerve defects, and then the collagen scaffolds combined with neurotrophic factors were used to promote axon regeneration and motor function recovery in rats with total transection of peripheral nerve. The experimental results show that the fusion of neurotrophic factors with collagen binding domain can specifically bind to collagen-based biomaterials, prevent rapid diffusion of factors in vivo, thus maintaining a more uniform release efficiency in vivo, which can be widely used in the study of motor ability recovery of peripheral nerve transection patients.

Bioinspired Scaffold Action Under the Extreme Physiological Conditions of Simulated Space Flights: Osteogenesis Enhancing Under Microgravity.

Prolonged exposure to microgravity (MG) during long-duration space flights is known to induce severe dysregulation of osteoblast functions connected to a significant bone loss, similar to the condition induced by osteoporosis. Hence, we here present MG as a promising model to challenge the effectiveness of new scaffolds designed for bone regeneration in counteracting bone loss. To this end, we carried out an integrative study aimed to evaluate, in the extreme condition of Random Positioning Machine-simulated MG, the osteoinductive potential of nanocrystalline magnesium-doped hydroxyapatite/type I collagen composite scaffold (MHA/Coll), that we previously demonstrated to be an excellent tool for bone tissue engineering. Initially, to test the osteoinductive properties of our bioinspired-scaffold, MHA/Coll structure was fully characterized under MG condition and compared to its static counterpart. Human bone marrow-derived mesenchymal stem cells were used to investigate the scaffold biocompatibility and ability to promote osteogenic differentiation after long-duration exposure to MG (up to 21 days). The results demonstrate that the nanostructure of MHA/Coll scaffold can alleviate MG-induced osteoblast dysfunction, promoting cell differentiation along the osteogenic lineage, with a consequent reduction in the expression of the surface markers CD29, CD44, and CD90. Moreover, these findings were corroborated by the ability of MHA/Coll to induce the expression of genes linked to osteogenesis, including alkaline phosphatase and osteocalcin. This study confirmed MHA/Coll capabilities in promoting osteogenesis even in extreme long-term condition of MG, suggesting MG as an effective challenging model to apply in future studies to validate the ability of advanced scaffolds to counteract bone loss, facilitating their application in translational Regenerative Medicine and Tissue Engineering.

Collagen, polycaprolactone and attapulgite composite scaffolds for in vivo bone repair in rabbit models

Although numerous materials have been explored as bone scaffolds, many of them are limited by their low osteoconductivity and high biodegradability. Therefore, new materials are desired to induce bone cell proliferation and facilitate bone formation. Attapulgite (ATP) is a hydrated silicate that exists in nature as a fibrillar clay mineral and is well known for its large specific surface area, high viscosity, and high absorption capacity, and therefore has the potential to be a new type of bone repair material due to its unique physicochemical properties. In this study, composite scaffolds composed of collagen/polycaprolactone/attapulgite (CPA) or collagen/polycaprolactone (CP) were fabricated through a salt-leaching method. The morphology, composition, microstructure, physical, and mechanical characteristics of the CPA and CP scaffolds were assessed. Cells from the mouse multipotent mesenchymal precursor cell line (D1 cells) were cocultured with the scaffolds, and cell adhesion, proliferation, and gene expression on the CPA and CP scaffolds were analyzed. Adult rabbits with radius defects were used to evaluate the performance of these scaffolds in repairing bone defects over 4–12 weeks. The experimental results showed that the cells demonstrated excellent attachment ability on the CPA scaffolds, as well as remarkable upregulation of the levels of osteoblastic markers such as Runx2, Osterix, collagen 1, osteopontin, and osteocalcin. Furthermore, results from radiography, micro-computed tomography, histological and immunohistochemical analysis demonstrated that abundant new bones were formed on the CPA scaffolds. Ultimately, these results demonstrated that CPA composite scaffolds show excellent potential in bone tissue engineering applications, with the capacity to be used as effective bone regeneration and repair scaffolds in clinical applications.

Multifarious Fabrication Approaches of Producing Aligned Collagen Scaffolds for Tissue Engineering Applications.

Aligned tissue architecture is a basic proviso for several organs and tissues like intervertebral discs, tendons, ligaments, muscles, and neurons, which comprises type-I collagen as an eminent extracellular matrix (ECM) protein. Exploiting type-I collagen for the biofabrication of aligned constructs via different approaches is becoming apparent, as it comprises a major fraction of connective tissue, exhibits abundance in ECM, and displays poor antigenicity and immunogenicity, along-with the ease of remodelling adaptability. Collagen hydrogels or composite scaffolds with uniaxial fibril alignment or unidirectional pore architecture having different sizes and densities are being fabricated using electrical, mechanical, and freeze-drying processes which are applicable for tissue engineering and regenerative purposes. This review focuses on several multifarious approaches employed to fabricate anisotropic structures of type-I collagen which influences fibril alignment, pore architecture, stiffness anisotropy, and enhanced mechanical strength and mimics the tissue native microenvironment ushering cell niches to proliferate and differentiate into tissue specific lineages.

The osteoinductive effect of nano-nacre particles on MC-3T3 E1 preosteoblast through controlled release of water soluble matrix and calciumions.

Prostheses and implants have been widely utilized in the orthopedic and dental fields. Nowadays, significant advances have been made in the structural and functional connection between living bone and prostheses, especially in the presence of compromised bone quantity/quality. Despite improvement in the treatment outcomes after augmentation, there are still challenges to meet the clinical demands due to limited materials available. In the current study, we investigated the effects of nano-nacre particles as an alternative material on stimulating bone cell differentiation and formation. Mouse osteoblastic cells (MC3T3-E1) were cultured on nano-nacre/type I collagen composite scaffold (NN-ICS) and type I collagen scaffold (ICS). Generated nano-nacre particles showed controlled release of protein and calcium for a period of 36 days. NN-ICS significantly contributed to the proliferation and differentiation of preosteoblasts compared to ICS controls. Our data showed that nano-nacre particles could serve as a candidate of bone substitution material, which potentially contributed to treatment outcomes in cases with compromised bone quality and/or quality.

Development and Characterization of Collagen Composite Scaffold for Cartilage Tissue Engineering

Development and Characterization of Collagen Composite Scaffold for Cartilage Tissue Engineering

Milieu for Endothelial Differentiation of Human Adipose-Derived Stem Cells

Human adipose-derived stem cells (hASCs) have been shown to differentiate down many lineages including endothelial lineage. We hypothesized that hASCs would more efficiently differentiate toward the endothelial lineage when formed as three-dimensional (3D) spheroids and with the addition of vascular endothelial growth factor (VEGF). Three conditions were tested: uncoated tissue culture polystyrene (TCPS) surfaces that induced a 2D monolayer formation; elastin-like polypeptide (ELP)-collagen composite hydrogel scaffolds that induced encapsulated 3D spheroid culture; and ELP-polyethyleneimine-coated TCPS surfaces that induced 3D spheroid formation in scaffold-free condition. Cells were exposed to endothelial differentiation medium containing no additional VEGF or 20 and 50 ng/mL of VEGF for 7 days and assayed for viability and endothelial differentiation markers. While endothelial differentiation media supported endothelial differentiation of hASCs, our 3D spheroid cultures augmented this differentiation and produced more von Willebrand factor than 2D cultures. Likewise, 3D cultures were able to uptake LDL, whereas the 2D cultures were not. Higher concentrations of VEGF further enhanced differentiation. Establishing angiogenesis is a key factor in regenerative medicine. Future studies aim to elucidate how to produce physiological changes such as neoangiogenesis and sprouting of vessels which may enhance the survival of regenerated tissues.

Dry versus hydrated collagen scaffolds: are dry states representative of hydrated states?

Collagen composite scaffolds have been used for a number of studies in tissue engineering. The hydration of such highly porous and hydrophilic structures may influence mechanical behaviour and porosity due to swelling. The differences in physical properties following hydration would represent a significant limiting factor for the seeding, growth and differentiation of cells in vitro and the overall applicability of such hydrophilic materials in vivo. Scaffolds based on collagen matrix, poly(DL-lactide) nanofibers, calcium phosphate particles and sodium hyaluronate with 8 different material compositions were characterised in the dry and hydrated states using X-ray microcomputed tomography, compression tests, hydraulic permeability measurement, degradation tests and infrared spectrometry. Hydration, simulating the conditions of cell seeding and cultivation up to 48 h and 576 h, was found to exert a minor effect on the morphological parameters and permeability. Conversely, hydration had a major statistically significant effect on the mechanical behaviour of all the tested scaffolds. The elastic modulus and compressive strength of all the scaffolds decreased by ~95%. The quantitative results provided confirm the importance of analysing scaffolds in the hydrated rather than the dry state since the former more precisely simulates the real environment for which such materials are designed.

Synthesis and characterization of collagen/PLGA biodegradable skin scaffold fibers.

The aim of this study is to investigate the applicability of poly(lactic-co-glycolic acid) (PLGA)/collagen composite scaffold for skin tissue engineering. PLGA and collagen were dissolved in HFIP as a common solvent and fibrous scaffolds were prepared by electrospinning method. The scaffolds were characterized by scanning electron microscopy (SEM), FTIR spectroscopy, mercury porosimetry, tensile strength, biocompatibility assays and Biodegradation. Cytotoxicity and cell adhesion were tested for two cell line groups, human dermal fibroblast (HDF) and human keratinocyte (HaCat). SEM images showed appropriate cell adhesion to the scaffold for both cell lines. MTT assays indicated that the cell viability of HDF cells increased with time, but the number of HaCat cells decreased after 14 days. The ultimate tensile strength was suitable for skin substitute application, but its elongation at break was rather low. For successful clinical application of the PLGA/collagen scaffold, some properties especially mechanical strain needs to be improved.

Ultralong hydroxyapatite nanowires/collagen scaffolds with hierarchical porous structure, enhanced mechanical properties and excellent cellular attachment

Abstract Hydroxyapatite/collagen (HAP/Col) composite porous scaffolds have been extensively investigated in the field of bone tissue engineering due to their similar chemical composition to that of natural bone and good biocompatibility. However, the relatively poor mechanical properties of the HAP/Col composite porous scaffolds limit their application in bone regeneration. In order to overcome these limitations, the hierarchical porous ultralong hydroxyapatite nanowires (UHANWs)/collagen (Col) composite scaffolds with excellent mechanical properties are synthesized by freeze-drying the suspension containing UHANWs and collagen. Compared with the HAP nanorods (HANRs), the highly flexible UHANWs can interweave with each other to construct a three-dimensional fabric-like structure in the collagen matrix of the UHANWs/Col scaffolds and significantly improve the mechanical properties of the scaffolds. The Young's modulus of the 70 wt% UHANWs/Col scaffold is about 4 times that of the pure collagen scaffold and 63 times that of the 70 wt% HANRs/Col scaffold. More importantly, in the absence of the collagen matrix, the pure UHANWs inorganic porous scaffold can be synthesized and exhibits relative good mechanical properties. However, the HANRs/Col scaffolds with HANRs weight fractions more than 70 wt% are easily cracked during the fabrication process. Furthermore, compared with the pure collagen and 70 wt% HANRs/Col scaffolds, the 70 wt% UHANWs/Col scaffold exhibits a superior biocompatibility and excellent ability to promote cell adhesion and spreading on the surface of the scaffold. In consideration of their outstanding mechanical properties and excellent cellular attachment performance, the as-prepared hierarchical porous UHANWs/Col scaffolds are promising for applications in various biomedical fields such as bone defect repair.

Construction and biocompatibility of a thin type I/II collagen composite scaffold

Articular cartilage injury is a common type of damage observed in clinical practice. A matrix-induced autologous chondrocyte implant was developed to repair articular cartilage as an advancement on the autologous chondrocyte implant procedure. Here, we establish a thin double layer of collagen as a novel and effective bioscaffold for the regeneration of cartilaginous lesions. We created a collagen membrane with double layers using a cover slip, a cover slip, and the collagen was then freeze-dried under vacuum. Carbodiimide as a crosslinking agent was used to obtain a relatively stable collagen construction. The thickness of the knee joint cartilage from grown rabbits was measured from a frozen section. Both type I and type II collagens were characterized using Sodium dodecylsulfate/polyacrylamide gel electrophoresis (SDS-PAGE) and ultraviolet absorption peaks. The aperture size of the scaffold was observed using a scanning electron microscope (SEM). The degradation of the scaffolds in vitro was tested through digestion using collagenase solution. The mechanical capacity of the scaffolds was assessed under dynamic compression. The influence of the scaffold on chondrocyte proliferation was assessed using the methyl thiazolyl tetrazolium (MTT) colourimetric assay and scanning electron microscopy. The frozen sections of the rabbit femoral condyle showed that the thickness of the weight-bearing area of the articular cartilage was less than 1mm. The results of the SDS-PAGE and ultraviolet absorption peaks of the collagens were in agreement with the standard photographs in the references. SEM showed that the aperture size of the cross-linked scaffold was 82.14±15.70μm. The in vitro degradation studies indicated that Carbodiimide cross-linking can effectively enhance the biostability of the scaffolds. The Carbodiimide cross-linking protocol resulted in a mean value for the samples that ranged from 8.72 to 15.95MPa for the compressive strength. The results of the MTT demonstrated that the scaffold had promoted chondrocyte proliferation and SEM observations showed that the scaffold was a good adhesive and growth material for chondrocytes. Thin type I/II collagen composite scaffold can meet the demands of cartilage tissue engineering and have good biocompatibility.