688 results about "Cell adhesion" patented technology

The invention relates to a high-activity recombinant XVII type collagen as well as a rapid preparation method and application thereof, and belongs to the technical field of bioengineering. The amino acid sequence of the high-activity recombinant XVII type collagen is as shown in SEQ ID NO. 1, and the nucleotide sequence of the high-activity recombinant XVII type collagen is as shown in SEQ ID NO. 2. The invention also provides a preparation method and application of the recombinant XVII type collagen. The high-activity recombinant XVII type collagen has good biological activity, has good effects of promoting cell adhesion, cell proliferation and cell migration, and can be compounded with other proteins to form raw materials of tissue engineering products, cosmetics, health care products or medicines. The high-activity recombinant XVII type collagen gene is integrated to different positions of a yeast genome through three different vectors, and the copy number of a target gene is increased, so that the expression quantity of the recombinant XVII type collagen is remarkably increased, and the yield of the high-activity recombinant XVII type collagen is effectively increased.

The invention discloses a recombinant humanized XVII type collagen as well as an expression vector and a genetically engineered bacterium thereof. Four functional fragments with high activity are obtained through screening, one of the fragments is optimized and then combined according to a certain sequence to obtain recombinant humanized XVII type collagen JYC1703, or one of the fragments is optimized and then combined according to a certain sequence, and GPP sequences are fused at an amino terminal and a carboxyl terminal respectively to obtain recombinant humanized XVII type collagen JYC1701 and JYC1702, and the recombinant humanized XVII type collagen JYC1701 and JYC1702 are combined according to a certain sequence to obtain the recombinant humanized XVII type collagen JYC1701 and JYC1702. After codon optimization, amplifying and cloning into an expression vector, transforming into pichia pastoris competent cells after enzyme digestion linearization, and screening high-copy recombinants through G418 resistance gradient, so as to obtain pichia pastoris genetically engineered bacteria for expressing recombinant humanized XVII type collagen. The recombinant humanized XVII type collagen JYC1701, JYC1702 and JYC1703 disclosed by the invention show excellent functions of promoting cell adhesion, cell proliferation and cell migration, and are suitable for preparing products such as skin care products or medical instruments and the like.

The invention discloses recombinant human XVII type collagen as well as a preparation method and application thereof. The recombinant human XVII type collagen is obtained by repeating an amino acid sequence from natural human XVII type collagen. The amino acid sequence of the recombinant human XVII type collagen is derived from natural human XVII type collagen, and the recombinant human XVII type collagen does not generate adverse immune response when being used for a human body. The recombinant human XVII type collagen can be secreted and expressed in a commercial yeast expression system, the expression quantity is greater than 10g / L, the preparation method is simple, and the high-yield recombinant human XVII type collagen can be obtained at low cost; in addition, the cell adhesion promoting activity of the recombinant human XVII type collagen is obviously superior to that of human collagen.

The invention provides humanized recombinant XVII collagen for promoting hair follicle stem cell repair and resisting aging as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The humanized recombinant XVII type collagen comprises polypeptide derived from the XVII type collagen or repeated protein formed by N times of repeated series connection of the polypeptide serving as a unit, n is an integer from 2 to 4; the polypeptide derived from the XVII type collagen comprises a polypeptide A and / or a polypeptide B; the amino acid sequence of the A polypeptide is as shown in SEQ ID NO: 1; and the amino acid sequence of the B polypeptide is as shown in SEQ ID NO: 2. The humanized recombinant XVII type collagen not only can be massively and stably recombined and expressed, but also has cell adhesion activity, cell proliferation promotion activity, cell migration promotion activity and excellent hair follicle repair and regeneration promotion activity, and has no sensitization side effect. The invention provides a new way for preparing collagen products with rich functions.

The invention relates to a recombinant A type humanized XVII type collagen as well as construction and application thereof, the protein is the humanized XVII type collagen which is composed of an intracellular region, a transmembrane region and an extracellular region, and the amino acid sequence is as shown in SEQ No. 1. The hair follicle repairing agent has the advantages of hair follicle repairing, hair regeneration, good biological activities of cell proliferation, cell adhesion and the like, and can be widely applied to the fields of medicines, medical instruments, biological materials, tissue engineering, cosmetics and the like. The gene sequence of the recombinant A type XVII collagen is 100% identical to the corresponding part of the gene sequence of the recombinant A type XVII collagen, so that the recombinant A type XVII collagen has good hydrophilicity and high activity, and has application potential in the fields of biomedical materials, beauty cosmetics, skin care products and the like.

The invention discloses recombinant human I-type collagen and application thereof. The amino acid sequence of the recombinant human I-type collagen is as shown in SEQ ID No: 2. The recombinant human I-type collagen has good cell proliferation promoting activity, cell migration promoting activity and cell adhesion promoting activity (superior to full-length recombinant human I-type collagen or commercially available human collagen) and is easy to produce. The recombinant human I-type collagen can be expected to have the effects of whitening, moisturizing, relieving and repairing, and can be applied to the fields of cosmetics, external medicines, skin care products, health care products, tissue engineering materials, medical instruments and the like.

The invention provides a recombinant XVII type collagen with a triple helix structure, and belongs to the technical field of biological proteins. The amino acid sequence of the collagen comprises one or more amino acid residue repetitive units, each repetitive unit comprises a sequence as shown in SEQ ID NO.1 or SEQ ID NO.2, the number of repetitions is n, and n is larger than 1 and smaller than or equal to 60. The recombinant XVII type collagen provided by the invention is simple in production process, relatively high in expression quantity, low in immunogenicity, high in safety, relatively good in stability of the triple-helix structure, and strong in supporting capacity for skin cells; efficacy experiments show that the collagen has higher cell adhesion, wound healing promotion and hair follicle repair promotion capability than XVII type collagen products in the market at present, and can be widely applied to the fields of food, health care products, cosmetics, medical instruments and the like.

The invention discloses humanized III-type collagen as well as a preparation method and application thereof, and belongs to the technical field of bioengineering. According to the method, firstly, on the basis of an original humanized III-type collagen gene, cell adhesion activity is taken as an evaluation index, a better cell adhesion sequence is screened out and spliced and integrated with the original collagen gene, and brand-new humanized III-type collagen is obtained. And constructing a recombinant expression vector of the humanized III type collagen containing the adhesion sequence by taking the probiotic EcN as a host cell, and introducing the recombinant expression vector into the EcN to obtain a strain of functional recombinant probiotic. Compared with a traditional escherichia coli engineering bacterium BL21 (DE3), the recombinant probiotics have good cell adhesion activity while efficiently expressing the humanized type III collagen, the bacterial endotoxin content of the recombinant humanized type III collagen obtained through purification meets the industrial standard, and the recombinant probiotics have good application prospects. The purification cost caused by industrial removal of endotoxin can be avoided, so that the production cost is reduced, and the method has a good application prospect.

The invention provides recombinant VIII type humanized collagen and application thereof, and the recombinant VIII type humanized collagen is of a triple helix structure, has good cell adhesion activity, does not generate immunological rejection and anaphylactic reaction when being applied to a human body, is a brand new human body synthetic biological material, and has good application prospects.

The invention relates to a protein-based phenylboronic acid ester dynamic crosslinking hydrogel material as well as a preparation method and application thereof. The preparation method comprises the following steps: dissolving a component A, namely a phenylboronic acid modified protein polymer derivative, in a biocompatible medium to obtain a solution A; dissolving a component B, namely a macromolecular derivative containing a vicinal diol group, into a biocompatible medium to obtain a solution B; and mixing the solution A and the solution B with a certain concentration for a certain time to obtain the protein-based phenylboronic acid ester dynamic crosslinking hydrogel material. The boric acid ester dynamic cross-linking bond belongs to reversible chemical cross-linking reaction, and the prepared protein-based hydrogel material has enough mechanical strength and cheap operability. The protein-based hydrogel material disclosed by the invention can provide the effects of cell adhesion, proliferation and differentiation promotion, tissue regeneration and the like, can load and slowly release beneficial components for promoting tissue repair, is an ideal tissue engineering scaffold material, and can be applied to defect regeneration and repair of soft tissues, cartilages, bones and the like.

Provided is a method for biosynthesis of a human structural material type IV collagen. Provided herein is a recombinant collagen, comprising sequences as shown in SEQ ID NO: 1 and the like. The recombinant collagen herein has good cell adhesion activity. The method herein relates to using the genetic engineering technology to produce the recombinant collagen, and overcomes defects in the prior art.

The present invention relates to a recombinant human type IV collagen and a preparation method therefor, which belong to the technical field of bioengineering. A recombinant human type IV collagen is designed on the basis of a partial sequence in an α2 chain of human type-IV collagen, and DNA encoding of the protein is ligated into an expression vector to construct a recombinant expression vector; then the recombinant expression vector is transformed into competent Escherichia coli cells, and a recombinant expression plasmid is extracted; after the plasmid is linearized, the linearized plasmid is transformed into an engineered strain HCPB-PPKEX2 to obtain a recombinant engineered strain; and the engineered strain is cultured, the expression of a target protein is induced, and the target protein is purified and identified. It is further demonstrated experimentally that the collagen can be completely cleaved by Kex2, Ste13 and CPB enzymes and efficiently secreted and expressed extracellularly, and the obtained sequence is completely consistent with a theoretical sequence. It is also verified experimentally that the recombinant collagen has a cell adhesion activity and a cell migration-promoting activity. The method avoids the risk of residual exogenous proteins, shortens the time and reduces the cost of the subsequent purification process, and thus has a relatively high safety and application value.

The invention relates to the technical field of bioengineering, in particular to a recombinant VII type collagen for inhibiting scars as well as a preparation method and application of the recombinant VII type collagen. The amino acid sequence of the recombinant VII type collagen is as shown in SEQ ID NO. 1. The recombinant VII type collagen provided by the invention has a better cell adhesion function, so that the recombinant VII type collagen can be applied to the fields of preparation of tissue materials, wound dressings and the like. Meanwhile, the VII type collagen does not contain any tag or exogenous amino acid sequence, is a completely humanized sequence, and does not generate immune response. The VII type collagen has a good adhesion effect, the adhesion effect is better than that of other recombinant collagen, the adhesion effect is equivalent to that of a common cell adhesive in the market, scars can be reduced by inhibiting activation of a TGF-beta pathway, and the VII type collagen can be developed into skin care products and medical instruments for promoting traceless wound repair and has a wide application prospect.

The present invention provides a yeast recombinant type XVII humanized collagen and its preparation method, in particular, a recombinant humanized collagen expressing the 15th helical region of type XVII collagen, a recombinant engineering bacterium and its preparation method, belonging to the technical fields of genetic engineering and synthetic biology. The present invention provides the full-length sequence of the 15th helical region of type XVII collagen, and mutants are obtained by mutating amino acid residues based on this sequence; for the first time, the present invention expresses the full-length sequence of the 15th helical region of type XVII collagen in Pichia pastoris, establishes a complete set of fermentation processes and purification processes, improves the protein expression level and reduces degradation, and through experiments, it is verified that the obtained protein has biological activities such as cell adhesion activity and cell migration promotion activity that reach or exceed those of natural human collagen, can achieve the purpose of real product application, and can be widely applied in the fields of medicine, medical devices, biological materials, tissue engineering, cosmetics, etc.

The invention relates to medical recombinant collagen repair gel and a preparation method thereof, and belongs to the technical field of recombinant collagen. Through the synergistic effect of the collagen, the chitosan, the polylactic acid, the nanocellulose, the vascular endothelial growth factor, the anti-inflammatory drug and the cell adhesion peptide, the tissue repair effect is remarkably improved. Through a gas foaming method, staged freezing and freeze drying, multistage pore size distribution is generated, gel with a porous structure is formed, and cell permeation and tissue regeneration are facilitated; in addition, the permeability and the drug loading capacity of the gel are improved, the cell adhesion peptides, the growth factors and the anti-inflammatory drugs are better adsorbed to the gel, functional slow release is achieved, and the repairing effect is further improved.

The invention discloses a bone repair material based on pH controlled release and a preparation method thereof. The bone repair material comprises the following raw materials: polycaprolactone, nano hydroxyapatite and nano magnesium oxide. The composite bone repair scaffold has the beneficial effects that the composite bone repair scaffold with excellent biocompatibility, excellent osteogenesis performance, excellent degradation performance and pH controlled release of magnesium ions is prepared by printing the bone repair material through a 3D printing technology, a temporary three-dimensional structure can be provided for cells, cell adhesion, proliferation and differentiation are promoted, growth factors are released at the same time, and the growth rate of the cells is increased. The formation of new bone tissues is guided; the composite bone repair scaffold can continuously and quantitatively release magnesium ions through pH value control, when osteoarthritis or bone tissue inflammation occurs at a bone defect part due to improper operation and rejection reaction, the magnesium ion release speed of the composite bone repair scaffold is increased, the anti-inflammatory effect is achieved, and the magnesium ions which are continuously released can promote bone tissue repair.

The invention discloses a recombinant human IV-type collagen as well as a preparation method and application thereof, and belongs to the field of bioengineering and protein engineering. According to the invention, three recombinant human IV type collagen new molecules are designed and screened on the basis of a part of sequences in a human IV type collagen alpha1 chain, soluble expression of a target protein is realized by using an escherichia coli system, and the recombinant collagen with the purity of more than 98% can be obtained through nickel column purification and acid regulation treatment. The recombinant collagen provided by the invention has good cell migration promotion activity and cell adhesion activity, and can be widely applied to the fields of tissue engineering, medical equipment, cosmetics, health care products and the like.

The invention discloses a collagen-based composite material and a preparation method thereof. The invention belongs to the technical field of composite materials. The collagen-based composite material comprises the following components in parts by weight: 50-60 parts of collagen; 10 to 20 parts of nano cellulose; 20 to 30 parts of a reinforcing component; 1-3 parts of a cross-linking agent; and 0.5-2 parts of a functional additive. By adding the reinforcing component, the mechanical strength of the collagen-based composite material is remarkably improved, and meanwhile, the non-toxic cross-linking agent and the natural reinforcing component are adopted, so that the material has good biocompatibility and does not generate toxic or adverse reaction on cells and tissues; according to the present invention, the porous three-dimensional network structure is provided, the pore size is 50-300 [mu] m, the porosity is more than or equal to 80%, the cell adhesion, the proliferation and the nutrient substance transmission are easily achieved, and the method is suitable for tissue engineering and wound repair; according to the collagen-based composite material disclosed by the invention, through reasonable raw material formula design, the collagen, the nano cellulose and the natural polysaccharide are compounded, the advantages of all the components are fully exerted, and the comprehensive performance of the composite material is further improved.

The invention relates to a method for rapidly detecting the cell adhesiveness of probiotics and application of the method. The method comprises the following steps: adding the probiotics into adherent culture cells, incubating, discarding liquid, digesting and collecting the cells by pancreatin, extracting total DNA (Deoxyribonucleic Acid), performing qPCR (Quantitative Polymerase Chain Reaction) amplification, and calculating the adhesion quantity of the probiotics on a single cell according to a Ct value. The invention provides a method for rapidly detecting the cell adhesion rate of probiotics by using a qPCR (quantitative polymerase chain reaction) technology, which comprises the following steps of: extracting total genome DNA (deoxyribonucleic acid) after the probiotics are adhered to cells on the basis of the gene expression difference of prokaryotic and eukaryotic cells, and detecting the specific gene expression level of the probiotics and the cells by using the qPCR technology. The cell adhesion ability of the probiotics is evaluated by calculating the ratio of the gene expression quantities of the two, and the method has the characteristics of high efficiency, rapidness, high accuracy and the like.

The invention discloses a method for fermenting recombinant humanized collagen by using escherichia coli and application of the recombinant humanized collagen, and belongs to the field of collagen. The amino acid sequence of the prepared recombinant humanized III-type collagen is shown as SEQ ID NO.4-7, the recombinant humanized III-type collagen is formed by connecting a plurality of repetitive units of natural collagen in series and has the advantages of being good in biocompatibility and high in safety, in addition, the nucleotide sequence for coding the recombinant collagen is designed according to the codon optimization theory, and the recombinant humanized III-type collagen has the advantages of being good in biocompatibility and high in safety. And carrying out fermentation expression on escherichia coli to obtain the recombinant humanized III-type collagen. The humanized III-type collagen can significantly improve the cell adhesion activity and proliferation activity, and can be widely applied to the fields of food, cosmetics, health care products, medical instruments and the like.

The invention provides recombinant IV type collagen and application thereof, and relates to the technical field of protein engineering. In the invention, an amino acid residue repetitive unit with good biological activity and stability in the human-derived IV-type collagen is found, and the amino acid residue repetitive unit is used as a core for repeating and constructing the recombinant IV-type collagen, so that the novel human-derived recombinant IV-type collagen with biological activity is obtained. The human-derived recombinant IV-type collagen provided by the invention is a small-molecular-weight protein, is easy to absorb by a human body, is good in safety, can remarkably promote secretion of collagen, also has the effects of promoting cell adhesion and cell migration, and has a good application prospect in the aspects of basement membrane repair and remodeling. A new functional component is provided for products for repairing and tightening the skin, resisting aging, resisting wrinkles, preserving moisture and the like.

The invention provides a methacrylic anhydride modified sclera extracellular matrix repair material and a curing method thereof, and belongs to the technical field of medical repair materials. The methacrylic anhydride modified sclera extracellular matrix repair material provided by the invention is prepared from the following raw materials: a methacrylic anhydride modified sclera extracellular matrix, methacrylated gelatin, a photoinitiator and a solvent. The sclera extracellular matrix modified by methacrylic anhydride is adopted, methacrylic anhydride contains methacrylic acid, double bonds can be introduced, the sclera extracellular matrix has the photo-initiation polymerization capacity, and in-situ gelling can be carried out at sclera and cornea damage positions under the action of a photoinitiator; the sclera extracellular matrix has good biocompatibility and histocompatibility, so that sclera matrix cells and corneal epithelial cells are adhered and proliferated, and the sutureless repair is realized; the methacrylated gelatin can provide a photo-crosslinking gelatin network under the action of a photoinitiator, so that cell adhesion and proliferation can be promoted, and repair is realized.

The invention discloses recombinant IV-type collagen as well as a preparation method and application thereof, and belongs to the technical field of bioengineering. The amino acid sequence of the recombinant IV type collagen comprises a plurality of repetitive units, and the repetitive units are shown as SEQ ID NO. 1. The recombinant IV type collagen fragment is derived from human IV type collagen, heterologous amino acid residues are not introduced, and rejection and sensitization risks are avoided; meanwhile, the hydrogel also has good biological activity, including cell proliferation promoting activity and cell adhesion promoting activity, and is good in biocompatibility. In addition, the preparation method of the recombinant IV-type collagen is simple and easy to operate, and the recombinant IV-type collagen with high purity and stability can be obtained through the fermentation and purification method. Therefore, the recombinant IV type collagen disclosed by the invention has a relatively good application prospect.

The invention discloses a preparation method of antibacterial and hemostatic medical sponge, and relates to the technical field of preparation of medical sponge. The preparation method of the antibacterial hemostatic medical sponge comprises the following steps: cleaning silkworm cocoons after removing impurities, and immersing the cleaned silkworm cocoons in a neutral protease solution to complete degumming so as to obtain fibroin fibers; the preparation method comprises the following steps: dissolving chitosan in an acetic acid solution, dissolving polyvinyl alcohol in deionized water, adding sodium alginate, adding a silk fibroin solution to form a composite solution, dropwise adding an AgNO3 solution, introducing CO2, maintaining the pressure, quickly releasing the pressure, foaming, injecting into a mold, carrying out gradient freezing to form a multistage-aperture sponge matrix, immersing in a genipin ethanol solution, and transferring into a 5% CaC 12 solution for soaking. The silk fibroin provides a biocompatible substrate and promotes cell adhesion, the sponge is constructed into a hierarchical pore structure with a compact surface and a loose interior, and liquid absorption and blood coagulation factor enrichment are accelerated.

The invention relates to the technical field of medical instruments, in particular to a multi-layer ultra-light hernia patch which comprises a large-mesh electrostatic spinning tissue ingrowth layer, a mesh woven base material layer and an anti-adhesion layer which are sequentially connected in a stacked mode. Wherein after the large-mesh electrostatic spinning tissue ingrowth layer is implanted into a human body, a cell microenvironment can be better simulated, cell adhesion, differentiation and migration are facilitated, uniform distribution of newborn cells in the fiber direction of an electrospinning membrane is promoted, granulation tissues uniformly ingrow in a defect part, and the electrospinning fiber membrane and the tissues are cooperatively repaired; secondly, the middle layer can provide supporting performance and mechanical performance of the patch in the early stage for the patch, the defects of the electrospinning fiber membrane in the supporting performance and the mechanical performance are overcome, the long-term implantation amount of the patch after later degradation is greatly reduced, and the foreign body sensation is obviously reduced; the anti-adhesion layer also adopts the electrostatic spinning technology, and the anti-adhesion effect is achieved by selecting materials and regulating and controlling the porosity, the aperture and the fiber diameter of the electrospun fiber membrane.

The invention discloses an intelligent integrated system for manufacturing a biological tissue engineering scaffold, relates to the technical field of biomedicine, and is used for improving the biocompatibility, mechanical strength and cell adhesion capacity of a biological scaffold. The plasma generator comprises a microwave generator, a precise electrode and an efficient gas flowing unit; the integrated manufacturing module comprises a 3D printing unit, a plasma enhanced chemical vapor deposition (PECVD) unit and a cooperative control unit; the monitoring and control system comprises an intelligent temperature and humidity sensor, a gas component monitor and an automatic control unit; the data management and optimization module comprises a biological safety evaluation unit, a personalized design and simulation unit, a material selection and management unit, a degradable material processing unit and a dynamic feedback and adjustment unit. According to the invention, low-temperature plasma treatment, 3D printing and PECVD technologies are fused, and a highly intelligent and integrated manufacturing platform is constructed.

The invention discloses an extracellular matrix fusion protein as well as a preparation method and application thereof, relates to the technical field of recombinant proteins, and aims to solve the technical problems that protein cosmetics in the market are relatively single in effect and difficult to meet application requirements. According to the extracellular matrix fusion protein, the nucleotide sequence of the extracellular matrix fusion protein is as shown in SEQ ID No.7. According to the extracellular matrix fusion protein with the nucleotide sequence shown as SEQ ID No.7, an in-vitro cell test proves that the fusion protein can remarkably promote cell adhesion, proliferation and cell migration, can act on a corium layer in a transdermal mode and remarkably promote collagen secretion, and due to the fact that recombinant protein belongs to biological protein, the fusion protein can be used for preparing the extracellular matrix fusion protein with the nucleotide sequence shown as SEQ ID No.7. Compared with common chemical cosmetics and hormone drugs, the composition has the advantages of easiness in decomposition, no residue and the like, and has a wider application prospect.

The invention discloses a preparation method and application of collagen microspheres, and belongs to the technical field of genetic engineering. Two histidines are respectively inserted into the N end and the C end of the collagen B-domain amino acid sequence, the high polymerization self-assembly of the collagen is promoted by utilizing the metal chelating effect to form a three-dimensional network structure, and a histidine is further inserted into the middle of the collagen, so that the three-dimensional morphology transformation of the collagen assembly is realized, and a loose and porous microsphere structure is formed. Experiments prove that by controlling the number and position of His residues in the collagen B-domain amino acid sequence expressed by microbial cells, the three-dimensional morphology of collagen can be controlled, and the collagen microspheres are formed; the prepared collagen microspheres can significantly promote cell adhesion, extension and migration.

The invention provides transdermal recombinant keratin as well as a preparation method and application thereof, and relates to the technical field of genetic engineering. The transdermal recombinant keratin comprises a recombinant keratin and a cell penetrating peptide; the transdermal recombinant keratin has cell adhesion activity and transdermal property. The transdermal recombinant keratin comprises any one amino acid sequence as shown in SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3 and SEQ ID NO.4 or consists of the amino acid sequence as shown in SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3 and SEQ ID NO.4. According to the transdermal recombinant keratin, the transdermal performance and the cell adhesion activity of keratin are remarkably improved, and the problem that traditional recombinant keratin is poor in skin permeability is solved.

The present invention belongs to the technical field of optimized coding gene, and discloses a recombinant human type I collagen, a preparation method thereof and an application thereof, and its amino acid sequence is shown as SEQ ID No.1. The recombinant type I collagen provided by the present invention has a high expression level, has better cell migration promotion, cell proliferation promotion and cell adhesion promotion properties, has high safety, and can be widely applied to fields such as medicine, medical devices, biological materials, tissue engineering, cosmetics and the like.